Objective To study genetic alterations in 9 STR loci and the Amelogenin locus in various tumor tissues. Methods twenty cancer tissues taken from 20 different unrelated individuals and their blood specimens were examined with Chelex-100 extraction of DNA, Profiler Plus PCR amplification and 310 Genetic Analyzer. Results All of the 10 STR loci exist genetic alterations. The genetic alterations occurred in 6out of 20 cases. The rate of genetic alteration was 30%. Six genetic alterations were found in one tumor tissue. Conclusion The forensic community has to take be cautious not to use the tumor tissue for personnel identification.

The key of Ankle-brachial index (ABI) measurement is the synchronous measurement of four limbs' systolic blood pressures. In this paper is analyzed the inadequacy of the modern blood pressure measurement technologies used in the ABI measurement process. Special emphasis is laid on the principles and characteristics of the double-layer cuffs technology. The research orientation, the existing problems, and the way toward improvement are discussed.
Ankle ; blood supply ; Ankle Brachial Index ; instrumentation ; methods ; Automation ; Blood Pressure ; physiology ; Blood Pressure Determination ; methods ; Brachial Artery ; physiology ; Humans ; Peripheral Vascular Diseases ; diagnosis ; physiopathology ; Regional Blood Flow

Objective To evaluate the clinical significance of high frequency ultrasonography in the diagnosis of annular pancreas in neonates.Methods Ultrasound results,clinical data,operation results and complicating deformity of 19 neonates were reviewed retrospectively.The digestive tracts of 19 neonates,including stomach,duodenum,jejunoileum and colon,were examined with 8-12 MHz linear transducer before surgery.Results Of 19 neonates with annular pancreas,17 cases were diagnosed with ultrasound,the diagnostic rate was 89％.The pancreas tissue encircling the descending duodenum was directly displayed as a direct sign of ultrasonography and the expansion of the proximal end of the duodenum was presented as an indirect sign of ultrasonography in 17 neonates.There were 3 neonates with annular pancreas complicating intestine malrotation and 2 neonates with annular pancreas complicating duodenal membraneous stenosis.One neonate was misdiagnosised and the other was missed.Conclusions High frequency ultrasonography plays an important role in diagnosing annular pancreas and other digestive deformity in neonates.It can be used as the first choice for the neonatal annular pancreas.

BACKGROUND: Revascularization is a challenge for the tissue-engineered bone carrying cells after implanted into human body. Previous studies have found that tanshinol can improve the functions of endothelial progenitor cells and exert vascular protective effects.OBJECTIVE: To prepare the β-calcium phosphate (β-TCP) scaffold with tanshinol coating, and to observe its cytocompatibility.METHODS: The β-TCP scaffolds coated with 10-7, 10-6 and 10-5 mol of tanshinol were constructed by negative pressure absorption method. The distribution of tanshinol coating on the scaffold was observed using scanning electron microscopy,and the inner ingredients were analyzed by infrared spectrum. Human endothelial progenitor cells (hEPCs) were cultured in the extracts of β-TCP and β-TCP scaffolds with 10-7, 10-6, 10-5 mol of tanshinol coatings, respectively. The cell proliferation was detected at 2, 4, 6, 8 and 10 days of culture; the levels of nitric oxide and vascular endothelial growth factor in the supernatants were detected at 1, 7 and 14 days of culture; the lumen formation on the matrigel was observed after 14-day culture. hEPCs were respectively seeded onto the β-TCP and β-TCP scaffolds with different dosages of tanshinol coating,and then the cell growth was observed under scanning electron microscope at 7 days.RESULTS AND CONCLUSION: The tanshinol coating evenly distributed on the inner surface of the pores, and its crystalline structure became dense with dosage increasing. Infrared spectrum analysis revealed no changes in the characteristic absorption peak of tanshinol and TCP in the scaffold. The β-TCP scaffolds with tanshinol coating could promote the proliferation of hEPCs, especially the scaffolds with 10-6 and 10-5 mol tanshinol coating. Compared with the β-TCP scaffold, the scaffolds with 10-6 and 10-5 mol tanshinol coating significantly upregulated the nitric oxide level at 14 days of culture, and significantly increased the level of vascular endothelial growth factor at 7 and 14 days of culture (P <0.05). Although it could be found in all β-TCP scaffolds with tanshinol coating, the lumen formation was the maturest in the scaffold with 10-5 mol tanshinol coating. These results suggest the β-TCP scaffolds with tanshinol coating can promote the proliferation and endothelial differentiation of hEPCs, and hold a good cytocompatibility.

Objective To investigate the characteristics of the bone marrow mesenchymal stem cell(MSC)in children with aplastic anemia(AA)in vitro,and the expressions of tumor necrosis factor -α-induced protein -8 -like 2(TIPE2)in the bone marrow,and the correlation between the level of TIPE2 mRNA with γ-interferon(IFN -γ)and IL -6 in AA patients.Methods Bone marrow samples were collected from 1 8 children with AA(AA group)and 8 children with bone injury (control group)who were hospitalized in Jinan Children′s Hospital from January 201 2 to June 201 5.MSC were isolated and cultured.The morphology of MSC was observed and immune phenotype was detected.The TIPE2 mRNA was detected by using real -time fluorescence quantitative PCR,and the levels of IFN -γand IL -6 were detected by using enzyme linked immunosorbent assay.Results Different sizes had been presented in the primi-tive MSC of AA patients,but the third passage MSC until 80% confluence had manifested the uniform convergence with long spindle and swirl distribution.In the sixth passage,cells showed degenerative change.The primitive and first pa-ssage MSC in patients with AA was longer than that in the controls.CD73 ,CD1 05 ,CD44 and CD90 were expressed in MSC,while CD34 ,CD45 ,CD271 expressed rarely.The level of TIPE2 mRNA in AA patients (5.29 ±1 .56)was obviously lower than that of the control group(8.68 ±2.00),and the difference was significant(t =-4.48,P <0.01 ).The con-centration of IFN -γ[(5.48 ±1 .97)ng/L]and IL -6[(5.43 ±1 .92)ng/L]in AA patients were higher than those of the control group[(3.40 ±1 .24)ng/L,(3.79 ±0.92)ng/L],and the differences were significant (t =2.70, 2.26,all P <0.05).The level of TIPE2 mRNA in AA patients was negatively related with IFN -γand IL -6(r =-0.838,-0.658,all P <0.05),but there was no significant correlation between them in the control group (all P >0.05).Conclusions The proliferation of MSC is significantly reduced in patients with AA.TIPE2,as an important role to stabilize the immune system,plays an important role in the occurrence of AA by its low expression and up -regula-ting the expression of inflammatory factors.

Objective To identify the genetic cause of a case of congenital hypotrichosis by a nextgeneration sequencing technology.Methods A 9-year and 3-month-old girl presented with few villous hairs at birth,which grew slowly.Skin examination showed sparse,thin,soft,woolly and light-yellow hairs,small amount of hairs on the top of the head and a less amount of hairs around the head,hairline recession and broadened forehead.No abnormality was found by ophthalmic examination.No similar aberrant phenotype was observed in the patient's parents or her younger sister.Her parents were non-consanguineous marriage.Peripheral venous blood samples were obtained from the patient,her mother and younger sister.Genomic DNA was extracted and then analyzed by a next-generation sequencing technology.The suspected pathogenic mutations were validated by Sanger sequencing and subjected to bioinformatics analysis.Results Two mutations were identified in the CDH3 gene in the patient,including a c.1057G ＞ T (p.D353Y) heterozygous mutation in exon 5 and a c.1767delC (p.I589Ifs) heterozygous mutation in exon 10.They were both novel mutations,and their pathogenicity was predicted by softwares.Sanger sequencing indicated that the c.1057G ＞ T (p.D353Y) heterozygous mutation was inherited from the patient's mother,and gene transfer analysis revealed that the c.1767delC (p.I589Ifs) heterozygous mutation was inherited from the patient's father.Conclusion The c.1057G ＞ T (p.D353Y) and c.1767delC (p.I589Ifs)heterozygous mutations may cause hypotrichosis and juvenile macular dystrophy in the patient,so careful observation and comprehensive ophthalmic examination should be performed on time for early diagnosis and treatment of eye symptoms.

OBJECTIVETo explore the genetic cause and clinical features of a neonate with bullous congenital ichthyosiform erythroderma.

METHODSThe patient was examined thoroughly. Following the extraction of genomic DNA, next generation sequencing was performed to analyze the genetic cause.

RESULTSThe patient manifested generalized erythema, blistering, and extensive exfoliation of the skin. A heterozygous missence 482T>G mutation was found in the first exon of KRT10 gene, which led to a p.L161W alteration in its protein product.

CONCLUSIONThe de novo mutation of the KRT10 gene probably underlies the disease in the child.

OBJECTIVETo delineate the clinical and genetic characteristics of a girl featuring motor retardation, language retardation and regression, and light persisting diarrhea.

METHODSThe patient was clinically examined and tested by tandem mass spectrometry and next generation sequencing.

RESULTSThe proband could not stand and walk alone, and had light persisting diarrhea. She manifested language development retardation and regression. Laboratory tests were all normal, but the screening of metabolic disorders for urine and blood showed deficiency of short chain coenzyme A dehydrogenase due to elevated ethylmalonic acid and butyryl carnitine. By next generation sequencing, two compound heterozygous mutations of the ETHE1 gene, c.2T>A and c.488G>A, were discovered in the proband, which were respectively inherited from her father and mother. Bioinformatics analysis predicted both mutations to be pathogenic. The patient was diagnosed with ethylmalonic encephalopathy. Vitamin B1, B2, Coenzyme Q10, and L-carnitine were prescribed. The patient deteriorated and required liver transplantation at 4-year-1-month.

CONCLUSIONBased on the clinical and genetic analysis, the proband was diagnosed with ethylmalonic encephalopathy caused by ETHE1 gene mutation. Next generation sequencing has provided a powerful tool for the diagnosis of such disorders.

OBJECTIVETo analyze the clinical features and genetic basis of a female neonate with muscle weakness, abnormal brain magnetic resonance imaging and elevated blood lactate.

METHODSThe patient was subjected to clinical and laboratory examination. Next generation sequencing was carried out for the patient and her relatives.

RESULTSThe proband was diagnosed as small for gestational age, with clinical features including muscle weakness, abnormal brain magnetic resonance imaging, increased blood lactate, and acidosis. By genetic testing, a de novo PDHA1 mutation c.1133G to A (p.R378H) was identified, which was known to be pathogenic. The patient was diagnosed with pyruvate dehydrogenase complex deficiency disease (PDCDD), for which vitamin B1, coenzyme Q10, and L-carnitine were prescribed, and a ketogenic diet was recommended. Follow-up at 4-month-7-day found that her blood lactic acid was reduced to normal but her muscle tone was still low.

CONCLUSIONThe proband was diagnosed as PDCDD caused by a PDHA1 missense mutation. NGS has provided a powerful tool for the diagnosis of such diseases.

Objective To study the molecular genetic mechanism of congenital central hypoventilation syndrome ( CCHS).Method The clinical data and molecular genetics results of CCHS diagnosed in neonatology department from 2014 to 2016 were analyzed retrospectively.The relationship between genotypes and clinical phenotypes in patients of CCHS was analyzed , and the diagnostic thinkings , follow－up and prognosis were summarized.Result A total of 4 infants with CCHS were included in this study.Among them, 2 were boys and the other 2 were girls.They were all full－term neonates without asphyxia at birth , but they soon sufferd from dyspnea and cyanosis , required assisted ventilation.One case had difficult defecation. All 4 cases had difficulty in weaning.The respiratory rhythm became weak developed apnea and carbon dioxide retention was detected in blood gas analysis.All the 4 cases died after withdrawal of treatment.The results of molecular genetic testing were as follows.There was a 38bp heterozygous deletion mutation in exon 3 of gene PHOX2B ( e.756＿776 del21bp).Three cases were found small fragment insertion in exon 3 of gene PHOX2B, which attributed to polyalanine repeat expansion mutations (PARMs).One case belonged to type 20/27 and another 2 cases belonged to type 20/26.Conclusion The main manifestation of CCHS in the neonatal period is ventilator dependant , which can combined with megacolon and atypical autonomic nerve disorder.According to the literature, more than 95%of CCHS are caused by the PHOX2B mutation. The symptom is severe when it got a non－PARMs mutation.It′s useful to make a definite diagnosis with genetic diagnosis results , which could be helpful for treating and predicting.Only effective respiratory support and standardized follow－up system can improve the quality of life in patients of CCHS.