BACKGROUND:Spermatogonial stem cells are a kind of adult stem cells, which have self-renewal and differentiation potential, and can be differentiated into specific cells in vitro, suggesting that the spermatogonial stem cells may be possibly differentiated into osteoblasts. But the related research has not been reported. OBJECTIVE:To observe the biological characterization and osteogenic process of mouse spermatogonial stem cells cultured in vitro. METHODS:Spermatogonial stem cells were obtained from the testicle of mice aged 15-20 days, and were cultured on the feeder layer from bone marrow stroma cells in vitro. When cultured for 3 days, the cells were cultured in the conditioned medium (experimental group) and basic medium (control group). The cells proliferation capability and osteogenic property were examined by phase-contrast microscope, alkaline phosphatase activity and type I col agen immunofluorescence staining. RESULTS AND CONCLUSION:Spermatogonial stem cells proliferated faster in the experiment group than in the control group. cells grew rapidly in colony-like shape in the conditioned medium at 3-6 days, the three-dimensional feeling enhanced, cellmass and clusters continued to increase in size, the extracellular matrix was increased in number and the cytoplasmic bridge was not obvious. After culture for 15 days, cells in the two groups were positive for alkaline phosphatase staining that the cytoplasmic membrane was dyed black. Under the fluorescent microscope, green fluorescence was visible in the experimental group, suggesting the cells in the experimental group was positive for type I col agen, but negative in the control group, which is similar with the biological characteristics of osteoblasts. These findings indicate that spermatogonial stem cells possess the osteogenic capability under induction conditions, which are expected to provide seed cells for bone tissue engineering.

Objective Acute kidney injury (AKI) is a common complication after cardiac surgery,especially in elderly patients,and related with poor prognosis.Although much advances in therapies of AKI have been obtained,the prognosis of patients did not improved.In the absence of proven interventions,a reasonable strategy would be to identify modifiable risk factors for AKI.The objective of the present study was to explore modifiable risk factors of acute kidney injury after cardiac surgery with cardiopulmonary bypass in elderly patients.Methods Data from 457 consecutive elderly patients (age ≥60 years old)who underwent cardiac surgery with cardiopulmonary bypass in the Guangdong General Hospital between January 2007 and December 31,2009 were analyzed in this retrospective research.The primary outcome was AKI according to the serum creatinine criteria of the RIFLE (renal Risk,Injury,Failure,Loss of renal function and End-stage renal disease) classification as an increase in serum creatinine ＞ 50％ from baseline to peak value within the first seven postoperative days.The baseline serum creatinine was defined as the latest serum creatinine before cardiac surgery.Univariate anadysis was carried out for patients'demographics data and multivariate analysis by logistic regression was used to obtain the independent risk factors for AKI.Results Among 457 elderly patients,patients mean age was (65.22 ± 4.17) years and they comprised 253 (55.4％) men and 204 (44.6％) women.AKI occurred in 313 (68.5 ％) participants.Compared with patients without postoperative AKI,the media length of intensive care unit was longer in patients with postoperative AKI,4.0 (2.0-7.5) days versus 2.0 (1.0-3.0) days,respectively.In logistic regression model,malc (odds ratio[OR] 1.894,95％ confidence interval[CI] 1.136-3.157),age above 65 years (OR 2.391,95％ CI 1.381-4.142),hypertension (OR 2.286,95％ CI 1.249-4.184),estimated glomerular filtration rate less than 60 ml/min (OR 1.933,95％ CI 1.111-3.362),preoperative uric acid ＞ 450 μ mol/L (OR 2.938,95％ CI 1.633-5.285),use of angiotensin converting enzyme inhibitors/angiotensin receptor inhibitors (ACE1/ARB) before cardiac surgery (OR 2.196,95％ CI 1.283-3.759),use of ACEL/ARB after surgery (OR 0.329,95％ CI 0.156-0.691),use of diuretics (OR 0.149,95％ CI 0.068-0.326),time of cardiopulmonary bypass above 120 min (OR 5.228,95％ CI 3.023-9.041) and prolonged mechanical ventilation (OR 2.921,95％ CI 1.527-5.586) were independent factors of AKI after cardiac surgery with cardiopulmonary bypass.Conclusion Preoperative uric acid above 450μmol/L was a modifiable risk factor of AKI after cardiac surgery with cardiopulmonary bypass in elderly patients.Therapies aimed at mitigating high preoperative uric acid may offer protection against this complication.

Objective To investigate the inhibitory mechanism of emodin on lipopolysaccharide(LPS)-induced nitric oxide(NO)generation in rat peritoneal macrophages.Methods NO production and iNOS expression were measured through nitrite assay and Western blotting assay,respectively.NF-kB activity and nuclei P65 expression were estimated by dual-luciferase and Western blotting assay,respectively.Intracellular free Ca2+([Ca2+]i)was detected using the ratiometric fluorescent calcium indicator dye,Fura-2,and a microspectrofluorometer.PLC-γphosporylation was analyzed by Western blotting assay.Results First,emodin was found playing active roles in suppressing LPS-induced NF-kB activation in rat peritoneal macrophages.Second,emodin down-regulated transient[Ca2*]i and could increase in NF-kB upstream signal.Finally,emodin suppressed phosphorylation of PLC-γ by LPS stimulation in the upstream of[Ca2+]i.Conclusion Suppression of PLC-γ phosphorylation is involved in emodin inhibiting NO generation by LPS stimulation in rat peritoneal macrophages.

Objective To investigate the reference interval of platelet‐related parameters including mean platelet volume (MPV) ,platelet distribution width (PDW) ,platelet hematocrit (PCT) .Methods Used Sysmex XN‐9000 assembly line and PEN‐TRA 120 hematology analyzer to analysis the venous blood cells for 3 415 cases of 16 to 90 years old physical examination personnel in Mianyang Central Hospital ,the platelet‐related parameters results of 2 718 standard healthy population by screening were statis‐tically analyzed .Results Platelet‐related parameters 95% reference interval (2 .5% -97 .5% ) of XN assembly line :M PV 9 .9-14 . 7 fL ,PDW 11 .2% -24 .3% ,PCT 0 .15% -0 .30% ;Platelet‐related parameters of 95% reference interval of PENTRA 120:MPV 8 .4-12 .7 fL ,PDW 13 .8% -24 .8% ,PCT 0 .12% -0 .26% ;comparison between the instruments :MPV ,PDW ,PCT results showed significant differences (P<0 .05);comparison between men and women :PCT test results of XN assembly line were differ‐ent (P<0 .05) ,MPV ,PDW ,PCT results of PENTRA 120 were different (P<0 .05);comparison between age groups :PCT result of all ages between men and women were different in XN assembly line (P<0 .05) ,in PENTRA 120 ,MPV ,PDW ,PCT results be‐tween ages were different in men (P<0 .05) ,PCT results between ages were different in women (P<0 .05) .Conclusion Each la‐boratory should establish the laboratory supporting biological reference interval of platelet‐related parameters based on different in‐struments ;in practical applications ,should set the reference interval for different gender or age .

This study is to investigate the effects of indirubin on ATP-induced immune responses of macrophages. For this, neutral red dye uptake method was used to test phagocytosis, MTT assay was used for measuring cell death, and reactive oxygen species (ROS) was tested with fluorescent probe DHE. The data showed that extracellular ATP attenuated phagocytosis, induced cell death and increased ROS production, and these effects were restored by pre-treating with indirubin. This result suggested that indirubin blockade the effects of ATP on macrophages, because extracellular ATP-induced effects are dependent on P2 receptors, in particular P2X7 receptors. Furthermore, the effects of indirubin on the activation of P2 receptors were tested, in particular P2X7 receptors. The data showed that indirubin significantly decreased ATP-induced, P2 receptors mediated intracellular Ca2+ concentration ([Ca2+]i) rise and inhibited P2X7 receptor-based ethidium bromide (EB) dye uptake. These results suggested the inhibitory effects of indirubin on the activation of P2X7 receptors, which may underlying the effects on ATP induced ROS production, phagocytosis attenuation and cell death of macrophages.

ObjectiveTo explore the relations between the alkaline phosphatase(ALP) and the type of cerebral palsy. MethodsThe ALP level of 40 children with different types of cerebral palsy were examined with the Olympus AU-600 apparatus. ResultsThe ALP value of the children with spastic cerebral palsy is normal. The ALP value of the children with athetoid cerebral palsy is higher than spastic cerebral palsy(P<0.001).Conclusions The ALP value can help us to identify the type of cerebral palsy.

Breast cancer susceptibility gene 1(BRCA1) plays an important role in breast cancer development and progression. BRCA1 encodes a 1863-amino acid protein with two BRCA1 C-terminal (BRCT) domains at its C-terminus, BRCT1 and BRCT2. Many cancer-predisposing mutations are located in the BRCT domains, which have been shown to induce chromatin unfolding by use of an approach that allows visualization of large-scale chromatin structure through lac repressor/lac operator recognition. To map the important region of BRCT domain (amino acid residues 1642-1736), six deletion mutant constructs were made. The chromatin structure assay showed that amino acid residues 1691-1721 are involved in the induction of chromatin unfolding. To further localize the critical amino acid residues, ten alanine scanning mutant constructs were made. The chromatin structure assay demonstrated that the 1707IAGGK1711 region is critical for the chromatin unfolding activity. Based on the mapped important region, Blast analysis identified a novel homologous protein. Mapping of the BRCT1 domain may aid in the presymptomatic risk assessment and provide a valuable tool for further study on the BRCT1 structure and function.
BRCA1 Protein ; chemistry ; physiology ; Base Sequence ; Chromatin ; chemistry ; Cloning, Molecular ; Female ; Genes, BRCA1 ; Humans ; Molecular Sequence Data ; Mutation ; Protein Folding ; Structure-Activity Relationship

ObjectiveTo identify mutations ofGATA4 andGATA6 genes in children with isolated congenital atrial septal defect (ASD).Methods From November 2012 to November 2013, 101 patients with ASD (99 unrelated patients and one twin) who were submitted to catheter-based intervention and 100 ethnicity-matched children without congenital heart disease, blood disorders and chromosomal abnormalities were enrolled. The blood was collected. The coding regions and lfanking regions of theGATA4 andGATA6 genes were ampliifed by polymerase chain reaction and sequenced using the dideoxvnucleotide chain termination technique, and then compared with the normal sequence in the Genbank.Results Two novel heterozygous missense GATA6mutations, c. G145A and c. G151A, were identiifed in 2 unrelated ASD patients, which were not present in the controls. These two mutations predicted the conversion of glycine into serine at amino acid residue 49 (G49S) and glutamate into lysine at amino acid residue 52 (K52E). A heterozygous missenseGATA6 mutation c.43 G>C, which caused a conversion from glycine to arginine, was found in 9 ASD patients and 7 controls. A single nucleotide polymorphism c.99G>T, which did not cause amino acid conversion inGATA4 gene, was found.ConclusionsGATA6 gene is an important transcription factor in heart development. The mutation ofGATA6 gene may cause the change of its transcriptional activity, and lead to ASD.

Child ; Child, Preschool ; Female ; Follicle Stimulating Hormone ; blood ; Gonadotropins ; blood ; Humans ; Infant ; Infant, Newborn ; Male ; Sex Factors ; Time Factors

OBJECTIVETo investigate the frequency and type of PHEX gene mutations in children with X-linked hypophosphatemic rickets (XLH), the possible presence of mutational hot spots, and the relationship between genotype and clinical phenotype.

METHODSClinical data of 10 children with XLH was retrospectively reviewed. The relationship between gene mutation type and severity of XLH was evaluated.

RESULTSPHEX gene mutations were detected in all 10 children with XLH, including 6 cases of missense mutation, 2 cases of splice site mutation, 1 case of frameshift mutation, and 1 case of nonsense mutation. Two new mutations, c.2048T>C and IVS14+1delAG, were found. The type of PHEX gene mutation was not associated with the degree of short stature and leg deformity (P=0.571 and 0.467), and the mutation site was also not associated with the degree of short stature and leg deformity (P=0.400 and 1.000).

CONCLUSIONSMissense mutation is the most common type of PHEX gene mutation in children with XLH, and c.2048T>C and IVS14+1delAG are two new PHEX gene mutations. The type and site of PHEX gene mutation are not associated with the severity of XLH.

Adolescent ; Child ; Child, Preschool ; Familial Hypophosphatemic Rickets ; genetics ; Female ; Humans ; Infant ; Male ; Mutation ; PHEX Phosphate Regulating Neutral Endopeptidase ; genetics ; Retrospective Studies