Stem cell growth factor (SCGF) is an early-acting hematopoitic cytokine that has two isoforms including hSCGF with full length molecules and hSCGFbeta, 78 amino acids of which lost in the conserved calcium-dependent carbohydrate-recognition domain (CRD). It has been demonstrated that hSCGFbeta is strictly species-specific in regulating he-matopoiesis. This study was aimed to explore whether human SCGF can exert synergistic stimulatory effect on heterogenous murine CFU-GM progenitor. Firstly, hSCGF cDNA was amplified from human fetal liver cDNA library by using two-step PCR. The hSCGF mature peptide coding sequence was subsequently placed at downstream of glutathione S-transferase (GST) sequence in GST gene fusion expression vector. The results indicated that there existed an additional 60 kD protein compared with mock BL21 when the cells hosting recombinant plasmid were induced with IPTG at 37 degrees C. SDS-PAGE analysis demonstrated that the GST-hSCGF fusion protein mainly existed in insoluble form. When induced at low temperature (28 degrees C), the recombinant protein was mostly soluble. The GST-fusion recombinant protein was subsequently purified by using affinity chromatography. The clonogenic assay revealed that, unlike hSCGFbeta, hSCGF had the granulocyte/macrophage promoting activity (GPA) for murine bone marrow GM progenitor. It is concluded that, in contrast to human SCGFbeta, the intact molecular hSCGF may have no species specificity, implying that CRD domain in human SCGFbeta does not directly bind to corresponding SCGF receptor, but may have certain biological function.
Cloning, Molecular ; DNA, Complementary ; biosynthesis ; genetics ; isolation & purification ; Hematopoiesis ; genetics ; Humans ; Species Specificity ; Stem Cell Factor ; biosynthesis ; genetics ; isolation & purification

Bone marrow mesenchymal stem cells (MSCs) are multipotent tissue stem cells that can be induced in vitro to differentiate into a variety of cells such as osteoblasts, chondrocytes and adipocytes. MSCs are useful vehicles for both cell and gene therapy for a variety of diseases. Here, we injected human MSCs with enhanced green fluorescent protein (EGFP) into the striatum of Parkinson disease (PD) rat and examined their survival, migration, differentiation, and the behavior changes in PD rats, which will provide a theoretical foundation and technical method for clinic PD therapy by stem cells. The results showed that human bone marrow MSCs can survive in rat brain for a long time (exceeding 70 d). MSCs were found in multiple areas of the rat brain including the striatum, the corpus callosum, contralateral cortex and even the brain vascular wall. Immunocytochemical staining suggested that implanted cells expressed human neurofilament (NF), neuron-specific enolase (NSE) and glial fibrillary acid protein (GFAP). At the same time, remission in abnormal behavior of the PD rats appeared. Rotation scores decreased gradually from 8.86+/-2.09 r/min pre-transplantation to 4.87+/-2.06 r/min 90 d post-transplantation (statistic result showed P<0.05).
Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Cell Movement ; Corpus Striatum ; Green Fluorescent Proteins ; administration & dosage ; Humans ; Male ; Mesenchymal Stromal Cells ; cytology ; Parkinson Disease ; therapy ; Rats ; Rats, Wistar ; Stem Cell Transplantation ; methods ; Transplantation, Heterologous

BACKGROUNDWhile intra-articular injection of sinomenine hydrochloride has a therapeutic effect on osteoarthritis, it has a short half-life, and is thermolabile and photolabile. The aim of this research was to evaluate the sustained-release of sinomenine hydrochloride from an injectable sinomenine hydrochloride and sodium hyaluronate compound (CSSSI) and its therapeutic effect in a rabbit model of osteoarthritis following intra-articular injection.

METHODSAn injectable compound consisting of 1% sodium hyaluronate and 2.5% sinomenine hydrochloride was prepared and kept as the experiment group, and 2.5% sinomenine hydrochloride was prepared and kept as the control group. The cumulative mass release was measured at different time points in each group in vitro. Sixty-five male Zelanian rabbits were randomly divided into five groups: 15 (30 knees) each for the control, sodium hyaluronate, sinomenine hydrochloride, and CSSSI groups respectively, and five (10 knees) for the modeling group. Papain was injected into both knees of each rabbit for model establishment. Subsequently, 0.2 ml of the corresponding drugs was injected into the articular cavities of the remaining experiment groups, while the control group was treated with 0.2 ml normal saline. All groups were treated once a week for 4 weeks. Seven days after the last treatment, knees were anatomized to perform pathological observations and Mankin's evaluation of the synovium. Four groups were compared using the SPSS 13.0 software package.

RESULTSIn the in vitro sustained-release experiments, 90% of the drug was released in the experiment group 360 minutes following the injection. Comparison of the Mankin's evaluations of the four groups illustrated statistical discrepancies (P < 0.05). In further paired comparisons of the CSSSI group vs. modeling control/sodium hyaluronate/sinomenine hydrochloride groups, statistical significance was uniformly obtained. Moreover, sodium hyaluronate and sinomenine hydrochloride treatments showed significant improvement over the modeling control (P < 0.05), whereas sodium hyaluronate vs. sinomenine hydrochloride comparison failed to reach significance (P > 0.05).

CONCLUSIONSCSSSI has a sustained-release effect on sinomenine hydrochloride. Intra-articular injection of CSSSI was significantly better than the sole sodium hyaluronate or sinomenine hydrochloride for the treatment of osteoarthritis in a rabbit model.

Animals ; Hyaluronic Acid ; administration & dosage ; therapeutic use ; Injections, Intra-Articular ; Male ; Morphinans ; administration & dosage ; therapeutic use ; Osteoarthritis ; drug therapy ; Rabbits ; Random Allocation

BACKGROUNDArrhythmogenic right ventricular cardiomyopathy (ARVC) is one of the leading causes of sudden cardiac death. Recent studies have shown that ARVC, which is an inheritable genetic change, results from mutations in genes encoding desmosomal proteins. Plakophilin-2 is an important component of the desmosome. Because the full range of genetic variations related to ARVC is unknown and no related studies of the Chinese population have been reported, we aimed to investigate the genetic variation of plakophilin-2 in ARVC patients from the Southern Region of China.

METHODSGenomic DNA was isolated from peripheral blood samples of all 34 ARVC patients, who were screened through a clinical evaluation. They were used to detect variations in the sequences of the plakophilin-2 genes by polymerase chain reaction amplification in combination with direct sequencing.

RESULTSIn exon-1 of the plakophilin-2 gene, a deletion mutation (c.145_148 del GACA) was found in one family pedigree. The mutation was also found in exon-2, 4, and 11 of the plakophilin-2 gene. The QT interval dispersion of the ECG was considerably longer in the mutation group than in the non-mutation group of ARVC patients, and this result was statistically significant (P < 0.05).

CONCLUSIONWe discovered a plakophilin-2 mutation that prolongs the QT interval dispersion in the southern Chinese ARVC population.

Adult ; Arrhythmogenic Right Ventricular Dysplasia ; genetics ; Asian Continental Ancestry Group ; genetics ; Child ; China ; DNA Mutational Analysis ; Exons ; genetics ; Genetic Predisposition to Disease ; Humans ; Male ; Middle Aged ; Mutation ; Pedigree ; Plakophilins ; genetics

Objective To understand the main serotypes, antibiotic resistance profiles and molecular typing characteristics of Listeria monocytogenes（LM) isolated from foods in Shandong Province from 2013 to 2016. Methods The antibiotic sensitivity of LM was tested by broth microdilution method. The serotypes were determined by slide agglutination and PCR, and the molecular typing was carried out by pulsed field gel electrophoresis (PFGE) and multilocus sequence typing(MLST) . Results Most of 191 LM strains were sensitive to the eight antibiotics tested. Tetracycline resistance was most prevalent (15/191, 7.85%). There was no significant difference in the 8 antibiotic resistance monitored for 4 years (P=1.000). The serotype 1/2a, 1/2b and 1/2c accounted for 38.82% (66/170), 18.82% (32/170), and 42.36% (72/170), respectively. The patterns of SDSRZXDZ016, S2014L031 and SDSRZX030, totally accounted for 33.78%, were the dominant types. The main ST types were ST9, ST8 and ST121, which accounted for 81.18% (69/85). The clinical common types, ST3, ST7 and ST87 accounted for 8.23% (7/85), mainwhile new ST type was not found. Conclusion The LM strains isolated in Shandong Province from 2013 to 2016 were sensitive to most antibiotics, but some strains were resistant to tetracycline and erythromycin. The dominant serotypes were 1/2c and 1/2a. Serotype 4b, prone to outbreaks of listeriosis, was not found. The main PFGE types were SDSRZXDZ016, S2014L031 and SDSRZX030, which were continuously found from 2013 to 2016. The main ST types were ST8, ST9 and ST121. The clinical types, ST3, ST7 and ST87 were isolated from food and should be paid seriously attention to.

BACKGROUNDHuman epidermal growth factor 2 (HER2) is one of the most important prediction factors, but only 25% - 30% of breast cancer patients HER2 are positive. It is unknown whether there are other molecular markers that could be used to predict prognosis and recurrence in HER2 negative patients. This study investigated correlations of cyclin A2 and HER2 levels with clinical outcomes in 281 patients with invasive breast cancer in order to identify whether cyclin A2 can serve as a prognostic factor in HER2 negative patients.

METHODSImmunohistochemical staining was used to detect cyclin A2 and HER2 expression in 281 patients. Cyclin A2 and HER2 gene amplifications were analyzed using gene analysis and RT-PCR in 12 patients. Risk and survival estimates were analyzed using Log-rank, Kaplan-Meier, and Cox regression analysis; cyclin A2 and HER2 consistency with survival were analyzed using Kappa analysis.

RESULTSPatients with higher cyclin A2 and HER2 expressions had significantly shorter disease-free survival periods (P = 0.047 and P = 0.05, respectively). Kappa analysis performed that cyclin A2 and HER2 showed a low Kappa index (kappa = 0.37), allowing us to conclude that cyclin A2 and HER2 detect different pathologies. Gene analysis and RT-PCR showed that cyclin A2 was upregulated in patients with early relapse; the average increase was 3.69 - 2.74 fold.

CONCLUSIONSCyclin A2 and HER2 are associated with proliferation and high recurrence, particularly when combined. Cyclin A2 is easily detected by nuclear staining and might be a useful biomarker for recurrence risk in HER2 negative patients.

Adult ; Aged ; Aged, 80 and over ; Breast Neoplasms ; genetics ; metabolism ; Cyclin A2 ; genetics ; metabolism ; Female ; Humans ; Immunohistochemistry ; Middle Aged ; Multivariate Analysis ; Receptor, ErbB-2 ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

Objective To clone and express VP_2 gene from HBoV,and the expressed VP_2 protein was as the antigen in order to detect serum from children in Wenling area with lower respiratory tract infections.Methods The VP_2 gene was reeombined with the genome of Baculovirus,which infected the insect cell.The fusion protein with HA tag was applied to confirm the specificity of expressed protein.Furthermore,the recombinant protein was observed using electron microscopy.The 176 serum from children in Wenling area with lower respiratory tract infections was screened using Western blot.Results The expressed VP_2 protein was more than 60% in total proteins from insect cell,and MWt about 60 ×10~3.The virus-like particle(VLP)Was observed using electron microscopy,and size about 20nm.The 176 serum from children in wenling area with lower respiratory tract infections was screened using Western blot.The HBoV positive rate Was 2.28%(4/176).Conclusion The VP_2 protein from human bocavims was expressed in insect cell successfully.Through HA tag the VP_2 protein wag specific,and then the assay using SDS-PAGE with Western blot could detect and screen the antibody in serum from children with lower respiratory tract infections rapidly and accurately.

Objective To investigate pave a way for studying pathogenicty of HBoV.Methods Isolation and cell culture of HBoV by human bronchial epithelial cell line.which was founded in our laboratory.The morphology of the virus were primarily studied with a transmission electron microscope.In addition,transcript mRNA was detected in human bronchial epithelial cells,which was passaged and infected within HBoV,using the reverse-transcription polymerase chain reaction(RT-PCR).The amplified products nucleotide sequence of HBoV were sequencing and sequence analysis.Results Cytopathic effect(CPE)was observed after the aseptic residue of filtration of 2 case sputum specimens with HBoV,which was inoculated to the human bronchial epithelial cell line.The virus particles were observed in the cytoplasm,which were hexagonal or spherical in shape and 18-26 nm in diameter,bulk was 20 nm.cDNA amplieon obtained 295 bp fragment results of electrophoresis bands as same as NS1 region of the conserved matrix gene of publish sequence of HboV.PCR products nueleotide sequence of HboV were compared withcorresponding HboV GeneBank sequences.The comparison/alignment and construction of phylogenetic trees also point to an affiliation of the parvovirus to the species HBoV.Conclusion Isolation and identification of HBoV could be done in the human bronchial epithelial cell,and we found some characterizing CPE in the human bronchial epithehal cell after HBoV infection.The above studies pave a way for studying pathogenicty of human bocavirus.

OBJECTIVETo determine the prevalence of urolithiasis in young children fed infant formula (IF) contaminated with melamine, and the association between IF consumption and urolithiasis.

DESIGNA total of 2 733 children < or = 3 years of age on September 1, 2008 in two townships of Gansu Province, China were studied. Cases of urolithiasis were diagnosed by ultrasonography. Milk product consumption was determined by their caregivers. Remaining IF samples were tested for melamine and cyanuric acid.

RESULTSOf 2 733 eligible children in the two townships, 2 186 (80%) were enrolled in our study. Overall, 16.6% (362) of 2 186 children had urolithiasis. The prevalence was 24.6% in children exclusively fed Sanlu brand IF, 9.7% in those fed other IF, and 8.5% in those fed exclusively on other milk products. For children exclusively breast-fed, no urolithiasis was found (P < 0.05). The prevalence of urolithiasis was 11.4% in children fed 400 g of Sanlu IF, rising to 37.5% in children fed over 25 600 g. Of 48 Sanlu IF samples, 91.7% contained melamine (median = 1 800 ppm; range = 45-4 700) and 66.7% contained cyanuric acid (median = 1.2 ppm; range = 0.4-6.3). Melamine was also detected in 22.2% of 36 other brand IF (median = 27.5 ppm, range = 4-50).

CONCLUSIONSUrolithiasis was associated with melamine-contaminated IF. Although one product caused most morbidity, other milk products may have also contributed to the outbreak.

Child, Preschool ; Data Collection ; Food Contamination ; Humans ; Infant Food ; analysis ; Triazines ; toxicity ; Urolithiasis ; chemically induced

Humans ; Child ; Klinefelter Syndrome/complications* ; Lupus Erythematosus, Systemic/complications*