BACKGROUND:Col agen/bioactive glass composite materials possess excellent osteogenic potential and biocompatibility, but its application in bone tissue engineering is limited by mechanical property and degradation. OBJECTIVE:To construct col agen/bioactive glass/chitosan composite scaffolds with good mechanical property, anti-degradation ability and bone repair property.
METHODS:Bioactive glass/col agen composite scaffolds with chitosan as dispersant were prepared by lyophylization. Fourier transform infrared spectroscopy, scanning electron microscope, X-ray diffraction, and dynamic biomechanical testing were used to characterize the structure and properties of the composite scaffolds. RESULTS AND CONCLUSION:Results show that charge-attractions in pre-prepared bioactive glass/chitosan solution increased the homogeneity of bioactive glass dispersed in col agen gel and the compressive modulus and strength increased significantly due to the homogeneity and intermolecular interactions between chitosan and col agen. The enzymatic degradation rate and mineralization activity in the simulated body fluid were also lower because of a high degree of embedment of bioactive glass in col agen/chitosan matrix, and entanglement of col agen in chitosan at molecular level, which decreased the exposure of bioactive glass to the simulated body fluid, and col agen to enzyme solution.

Objective to investigate the efficacy of moxibustion treatment to rheumatoid arthritis(RA) patients and the impact to blood levels of rheumatoid factor(RF) ,C reactive protein(CRP) ,and interleukin-6(IL-6) .Methods RA patients were divided in-to treatment group and comfort group .The two groups were treated by moxibustion and sham-moxibustion for 2 courses ,respec-tievely .Results Overall clinical efficacy rate was higher in treatment group than in comfort group(P<0 .01) .The arthralgia ,ar-throcele ,tenderness and its degrees ,and morning stiffness time were significant improved in both groups(P<0 .05) ,and the indexes were better in treatment group than comfort group(P<0 .05) .The blood levels of RF ,IL-6 and CRP were reduced after treatment in both groups(P<0 .05) ,and which of treatment group reduced more than those of comfort group(P<0 .05) .Conclusion Moxi-bustion treatment can significantly improve clinical outcomes of RA and markedly reduce the blood levels of RF ,IL-6 and CRP of RA patients .

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.

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.

Infectious clone is a useful tool in exploring viral replication and pathogenesis. In order to prevent linear PCV2 cyclization, PCR mutagenesis was used to construct the first molecular clone (pSK-2PCV2) by ligating two copies of the complete PCV2 genome with the pBluescript SK (pSK) vector. In addition, pSK-PCV2 and ds-PCV2 were constructed. PK-15 cells were transfected with above three infectious clones. Indirect immunofluorescence assay (IFA) revealed that the virus antigen mainly localized in infected cell nucleolus and cytoplasm. PCV2 specific nucleotide fragment in cell culture was amplified by RT-PCR. Typical porcine circovirus particles with diameter about 17 nm were also observed by transmission electron microscope (TEM) in the infected cells. The rescued virus sequences from the cultures had 100% homology with the inserting PCV2 genome. The rescued virus shared similar properties with that of the parental virus. The study establishes a platform for further research on the virus molecular biology and pathogenicity.
Animals ; Cell Line ; Circoviridae Infections ; virology ; Circovirus ; genetics ; growth & development ; pathogenicity ; physiology ; Cloning, Molecular ; DNA, Viral ; genetics ; physiology ; Recombination, Genetic ; genetics ; Swine ; Transfection ; Virulence ; Virus Replication

OBJECTIVETo observe the repair effect of Schwann cells (SCs) modified by microgene pSVPoMcat on injured spinal cord in rats.

METHODSSemi-transection injury at the level of T(8) of spinal cord was made with cutting method on 120 Sprague Dawley (SD) rats. Then 40 rats implanted with SCs modified by microgene pSVPoMcat were taken as Group A, 40 rats implanted with simple SCs as Group B and the other 40 rats were taken as the control group (Group C). The functional recovery of the rats was observed through combined behavioral score (CBS) and cortical somatosensory evoked potential (CSEP), and the expression of the glial fibrillary acidic protein (GFAP) was measured with in situ hybridization and immunocytochemistry. At 3 months after operation, the rats were examined with magnetic resonance image (MRI), and the neurofilaments (NF) of the axons were stained with immunohistochemical method.

RESULTSGFAP expression in Group A was significantly lower than that of the other 2 groups. MRI showed that the spinal signals in the injured area recovered fundamentally in Group A, didn't recover in Group B and malacia focus was found in Group C, which was same as the results of NF staining. Wave amplitudes in incubation periods in Group A and Group B tended to recover. It recovered to the normal level in Group A, which was similar to the results of CBS.

CONCLUSIONSSCs modified by microgene pSVPoMcat can inhibit GFAP expression, improve the growth of the axons and the functional recovery of neurons after spinal cord injury.

Analysis of Variance ; Animals ; Evoked Potentials, Somatosensory ; Gene Transfer Techniques ; Genetic Therapy ; Glial Fibrillary Acidic Protein ; metabolism ; Immunohistochemistry ; In Situ Hybridization ; Magnetic Resonance Imaging ; Nerve Regeneration ; Rats ; Rats, Sprague-Dawley ; Schwann Cells ; metabolism ; transplantation ; Spinal Cord ; physiopathology ; Spinal Cord Injuries ; pathology ; physiopathology ; therapy

OBJECTIVETo study the curative effect of wilsonii injecta on severe head injury (SHI).

METHODSA total of 120 patients with SHI were divided randomly into 2 groups, the patients treated with conventional methods as Group A (n=60) and the patients treated with wilsonii injecta as Group B (n=60). The changes of neural function indexes were evaluated with Glasgow Coma Scale (GCS) before treatment and with Glasgow Outcome Scale (GOS) after treatment, simultaneously, the parameters of hemorrheological indexes (HI), brain electrical activity map (BEAM) and transcranial Doppler sonography (TCD) were observed before and after treatment.

RESULTSIn Group B, the clinical GCS, the HI, the BEAM and the prognosis GOS were improved much more than those in Group A. And the TCD parameters in Group B decreased, which had significant difference compared with that in Group A (P<0.01).

CONCLUSIONSWilsonii injecta can rapidly improve the injured p ersons' conscious states, the abnormal BEAM and the surviving quality. It suggests that the improvement of the HI is related to the relief of the vasospasm of the arterial blood vessels in the brain, which may be one of the important mechanisms of wilsonii injecta in improving the prognosis.

Adult ; Brain Injuries ; diagnostic imaging ; drug therapy ; Brain Mapping ; Female ; Follow-Up Studies ; Glasgow Coma Scale ; Glasgow Outcome Scale ; Humans ; Injury Severity Score ; Male ; Plant Extracts ; administration & dosage ; Probability ; Reference Values ; Treatment Outcome ; Ultrasonography, Doppler

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.

OBJECTIVE: To explore the genetic cause for a child with growth retardation by next generation sequencing (NGS). METHODS: Clinical data of the patient was collected. Peripheral venous blood samples were taken from the neonate and his parents. Targeted capturing and NGS were carried out to detect mutations of genes associated with inborn errors of metabolism. Suspected mutations were validated by Sanger sequencing. RESULTS: The 15-month-old female patient was admitted to hospital for growth retardation for 4 months. Hypomyelination was found upon cranium MRI. Genetic testing revealed two novel insertional mutations in the GLB1 gene in the patient, namely c.2006-2007insT and c.475-476 insGGTCC. CONCLUSION The c.2006-2007insT and c.475-476 insGGTCC mutations of the GLB1 gene probably underlie the GM1 gangliosidosis resulting in the growth retardation in the child.
Female ; Gangliosidosis, GM1 ; genetics ; Humans ; Infant ; Infant, Newborn ; Mutation ; Pedigree ; beta-Galactosidase ; genetics

OBJECTIVE: To explore the genetic basis for a neonate featuring hyperammonemia. METHODS: The patient was examined and tested by tandem mass spectrometry and next generation sequencing (NGS). Suspected mutations were confirmed by Sanger sequencing of the proband and her parents. Potential impact of the mutation was predicted with SIFT, PolyPhen-2 and MutationTaste software. RESULTS: Plasma ammonia and alanine were significantly increased in the proband, while serum citrulline was decreased. The neonate was found to harbor compound heterozygous mutations of the CPS1 gene [c.1631C>T(p.T544M) and c.1981G>T(p.G661C)], which were respectively inherited from her father and mother. CONCLUSION The carbamoyl phosphate synthetase I deficiency of the proband can probably be attributed to the mutations of the CPS1 gene. Above finding has expanded the spectrum of CPS1 mutations in association with carbamoyl phosphate synthetase I deficiency.
Carbamoyl-Phosphate Synthase (Ammonia) ; genetics ; Carbamoyl-Phosphate Synthase I Deficiency Disease ; genetics ; Female ; High-Throughput Nucleotide Sequencing ; Humans ; Hyperammonemia ; diagnosis ; genetics ; Infant, Newborn ; Mutation