OBJECTIVETo analyze the nature of the protein encoded by 8B7cDNA (1 835 bp) and to examine the localization of the protein in cells.

METHODSThe nature of the protein was analyzed using Blastn, Blastp, and TMpred. Expressions of 8B7 mRNA in tissues and cells were examined by Northern blotting. Recombinant expression vectors for localization test were constructed and transfected into COS-7 cells. Fluorescence emission in cells was examined upon a laser scan confocal microscope.

RESULTSThe protein encoded by 8B7cDNA was 363 amino acids long and contained spectrin repeats. It was completely homologous to the C-terminal 363 amino acids of Enaptin, Nasprin-1, Mynel, and Syne-1 proteins. It belonged to Spectrin super-family and was called 8B7 spectrin. Northern blotting revealed a 1.8 kb mRNA expression in human spleen and small intestine tissues. EGFP-8B7 fusion protein was observed to express on the nuclear membrane and in the cytoplasm in a reticular form in a localization assay on COS-7 cells. The position of fluorescence in COS-7 cells transfected with pEGFP-delta SR8B7 was similar to that in the cells transfected with pEGFP-8B7cDNA.

CONCLUSIONS8 B7 spectrin is a novel member of spectrin super-family. It localizes to the nuclear membrane and the cytoplasm in a reticular form in COS-7 cells. The localization is determined by the C-terminal KASH domain in 8B7 spectrin molecule.

Amino Acid Sequence ; Animals ; Blotting, Northern ; COS Cells ; Cercopithecus aethiops ; DNA, Complementary ; genetics ; Humans ; Plasmids ; genetics ; RNA, Messenger ; genetics ; Sequence Homology, Amino Acid ; Spectrin ; biosynthesis ; chemistry ; genetics ; Transfection

OBJECTIVETo study the value of bedside lung ultrasound in the diagnosis of neonatal pneumonia.

METHODSA total of 49 neonates who were admitted to the Neonatal Intensive Care Unit of Chengdu Women and Children's Central Hospital in March 2017 with respiratory symptoms as the chief complaint were enrolled. Bedside lung ultrasound was performed within 24 hours after admission. A retrospective analysis was performed for their clinical data and lung ultrasound findings. The value of bedside lung ultrasound in the diagnosis of neonatal pneumonia was evaluated.

RESULTSAccording to the gold standard for the diagnosis of neonatal pneumonia, of all 49 neonates, 44 were diagnosed with pneumonia. According to the criteria for the diagnosis of neonatal pneumonia based on lung ultrasound findings, 38 neonates were diagnosed with pneumonia. In the neonates with respiratory symptoms, lung ultrasound had a sensitivity of 86%, a specificity of 100%, a positive predictive value of 100%, and a negative predictive value of 45% in the diagnosis of neonatal pneumonia. Among the 44 cases of neonatal pneumonia diagnosed by the gold standard, the lung ultrasonic images showed B-lines in all 44 neonates (100%), 75% had pleural line abnormalities, 36% had patchy or local hypoechoic area in the lung, 27% had alveolar-interstitial syndrome, and 20% had air bronchogram.

CONCLUSIONSAs a new diagnostic technique in clinical practice, bedside lung ultrasound has a high sensitivity and specificity for the diagnosis of neonatal pneumonia and can thus be used as a tool for the diagnosis of this disease.

Female ; Humans ; Infant ; Infant, Newborn ; Infant, Newborn, Diseases ; diagnosis ; diagnostic imaging ; Lung ; diagnostic imaging ; Male ; Pneumonia ; diagnosis ; diagnostic imaging ; Retrospective Studies ; Ultrasonography ; methods

OBJECTIVEIn order to obtain functional genes, a normalized stems cDNA library was constructed from medicinal plant Dendrobium officinale.

METHODSMART (switching mechanism at 5' end of RNA transcript) cDNA synthesis combined with DSN (duplex-specific nuclease) normalization was applied to construct the normalized full-length cDNA library of D. officinale.

RESULTThe titer of cDNA library was about 1.3 x 10(6) cfu x mL(-1) and the average insertion size was about 1.5 kb with high recombination rate (93.9%). Random selected 163 positive clones were sequenced at single side. Bio-information analysis indicated that 147 from 150 high-quality unique sequences matched corresponding homologous proteins, and they participated in various biological processes based on GO (gene ontology). There were 8 clones with complete coding sequence, which presumed to be full-length genes.

CONCLUSIONThese results showed preliminarily that we successfully constructed a normalized full-length cDNA library of D. officinale which could be used to screen the functional genes related to metabolic pathways of medicinal ingredients.

Base Sequence ; Cloning, Molecular ; DNA, Complementary ; biosynthesis ; genetics ; Dendrobium ; genetics ; Gene Library ; Molecular Sequence Data ; Plants, Medicinal ; genetics ; RNA, Messenger ; genetics ; metabolism ; Sequence Analysis, DNA ; methods

This study was purposed to explore the changes of possible angiogenetic factors other than VEGF after inhibition of NHE1 and their related mechanisms. The K562 cells were treated by NHE1 specific inhibitor cariporide, the angiogenesis factors after inhibition of NHE1 were screened by using protein chip, the IL-8 expression level after cariporide treatment was detected by real-time quantitative PCR; the K562 cells with stable interference of NHE1 were constructed, the IL-8 expression level after interference of NHE1 was detected by real-time quantitative PCR; the p38 phosphorylation level in K562 cells treated with cariporide was detected by Western blot. After treatment of K562 cells with p38 inhibitor SB203580, the IL-8 expression level was decreased by real-time quantitative PCR. The results of protein chip showed that IL-8 expression decreased after cariporide treatment. Real-time quantitative PCR confirmed this inhibitory effect. The p38 phosphorylation level increased after cariporide treatment. The down-regulation of IL-8 expression induced by cariporide treatment was partially restored after K562 cells were treated with p38 inhibitor SB203580. It is concluded that the inhibition of NHE1 can inhibit IL-8 expression through up-regulation of p38 phosphorylation.
Cation Transport Proteins ; antagonists & inhibitors ; Down-Regulation ; Guanidines ; pharmacology ; Humans ; Imidazoles ; pharmacology ; Interleukin-8 ; metabolism ; K562 Cells ; Phosphorylation ; drug effects ; Pyridines ; pharmacology ; Sodium-Hydrogen Exchanger 1 ; Sodium-Hydrogen Exchangers ; antagonists & inhibitors ; Sulfones ; pharmacology ; p38 Mitogen-Activated Protein Kinases ; metabolism

OBJECTIVETo investigate the mechanism and the accelerating effect of rhEGF and rhbFGF on wound healing.

METHODSTwelve New Zealand rabbits with 72 incised wounds on ventral side of 24 ears were randomly divided into two therapeutic groups (rhEGF of 10 ug/cm(2) and rhbFGF of 100 AU/cm(2)) and a control group (1% silver sulfadiazine cream, SD-Ag). The general conditions of the wound healing was observed grossly. Biopsies were harvested at different time points for the pathomorphological examination, the electron microscopic examination, and for assessment of integrin beta1 mRNA expression by in situ hybridization.

RESULTSThe expressions of integrin beta 1 mRNA in two therapeutic groups were significantly higher than that of control group. The quality of the wound healing was improved in therapeutic group with its healing time shortened when compared with that in control group (P < 0.05). There was an obvious difference in the number of fibroblasts and capillary gemmules between the therapeutic and control groups (P < 0.05).

CONCLUSIONThe wound healing and quality could be improved by both rhEGF and rhbFGF, but rhbFGF seemed better to be employed during the early and middle stages of the wound repair for the growth of granulation tissue, while rhEGF should be applied at the late stage of wound repair to accelerate the re-epithelialization of the wound. Combined application of rhEGF with rhbFGF according to time effect could be more beneficial to the wound repair.

Animals ; Epidermal Growth Factor ; pharmacology ; Female ; Fibroblast Growth Factor 2 ; pharmacology ; Integrin beta1 ; genetics ; Male ; Microscopy, Electron ; RNA, Messenger ; analysis ; Rabbits ; Recombinant Proteins ; pharmacology ; Wound Healing ; drug effects

BACKGROUND:Epidermal growth factor is an auxiliary growth factor,but its effect on the growth of bone marrow mesenchymal stem cells (BMSCs) is uncertain.OBJECTIVE:To establish a mature method for isolation,extraction and identification of rat BMSCs,to investigate the effects of epidermal growth factor (EGF) on the proliferation and migration ability of BMSCs and to explore its potential mechanisms at the same time.METHODS:Rat BMSCs were isolated and purified using the improved whole bone marrow adherence method.After the cells were subcultured to the third generation,we detected the expression of cells surface antigens CD29,CD45 and CD90 by flow cytometry.BMSCs were further identified by osteogenic and adipogenic differentiation.Meanwhile,the effect of EGF on the proliferation of passage 3 BMSCs was measured by cell counting kit-8 and clonogenic assay.And the migration of P3 cells was verified by Transwell chamber.In addition,we detected the expression of proteins related to PI3K/Akt and nuclear factor-κB signaling pathways by western blot assay.RESULTS AND CONCLUSION:The primary BMSCs were polygonal and spindle-shaped,and then gradually appeared to be spindle-shaped.The results of flow cytometry demonstrated that the passage 3 cells were positive for CD29 and CD90,but negative for CD45.Furthermore,we successfully induced the osteogenic and adipogenic differentiation of BMSCs in vitro.Additionally,our data demonstrated that EGF promoted the proliferation and migration of passage 3 BMSCs.The relative expression levels of p-Akt and Bcl-2 of PI3K/Akt signaling pathway was up-regulated and the expression of Bax was down-regulated.At the same time,the relative expression level of phosphorylated p65 of nuclear factor-κB signaling pathway was up-regulated and the expression of phosphorylated inhibitor κB was down-regulated.Moreover,the downstream protein of matrix metalloproteinase-9 was up-regulated.Those proteins were related to the migaration of BMSCs.In summary,our results suggest that EGF could promote the proliferation and migration of BMSCs.

Objective To investigate the changes of matrix Metalloproteinases-9 (MMP-9) content in the cerebrospinal fluid of patients with brain injury and its clinical significance.Methods Sixty cerebrospinal fluid (CSF) specimens from patients with traumatic brain injury were chosen in our study as experimental group,and other CSF samples from 35 healthy subjects were selected as control group.The content of MMP-9 in cerebrospinal fluid was measured by enzyme-linked immunosorbent assay (ELISA) at 24 h and 72 h,1 and 2 weeks,respectively; and the comparative study was performed.Results The MMP-9 content in all CSF samples from the experimental group was significantly higher than that in the healthy control group (P＜0.05).Conclusion MMP-9 content in CSF of patients with brain injury increases obviously; MMP-9 in CSF may contribute to the severity of traumatic brain injury determination and guide the treatment strategies,which is worth for further study.

We investigated epidemiological characteristic of scrub typhus in Yongshan County,Yunnan Province,China. The serum samples were collected from the patients with fever for detecting the antibody against Orientia tsutsugamsushi (Ot) by colloidal gold immunoassay assay.Rat traps were used to capture rodents.The spleen tissues of the captured rodents were detected by nested-polymerase chain reaction for rickettsia groEL segment.The groEL segments were sequenced and analyzed the homology with the other known sequences.Thirty-four scrub typhus cases were found in Yongshan County,Yunnan Prov-ince from May 2015 to October 2015.Among them,21 cases were confirmed by laboratory tests and 13 cases were clinical di-agnosis diseases.Of these patients,32.35% of the cases occurred in June.The 32.35% were in the group of the 40-49 year-old,and 79.41% were farmers,94.12% exhibited eschar or skin ulcer(31.25% were observed in groin of these cases),and rash developed in 50%.In 39 spleen tissue samples of Rattus flavipectus,9 samples showed positive for groEL gene Ot,but gro-EL gene of Typhus group rickettsia and spotted fever group rickettsia were negative.Sequence analysis showed that YSP30 was closely related to some Saitama related strains of Ot,such as HSB1,FAR1 and UAP4,while the other 8 strains were closely related to some Karp related strains of Ot,such as UT213,UT221 and SH205.It was confirmed that the Yongshan County was the natural foci of scrub typhus by the serological and molecular biological detections.There are Karp and Saitama genotype related Ots in the natural foci.

Objective:To study the clinical value of N-terminal pro-B-type natriuretic peptide (NTproBNP) predicting the risk of bronchopulmonary dysplasia (BPD) in very/extremely low birth weight infants (VLBWI/ELBWI).Methods:From June 2017 to December 2019, VLBWI/ELBWI admitted to neonatal department in our hospital were enrolled in this non-interventional prospective study. According to the occurrence of BPD, the infants were assigned into BPD group and non-BPD group. Infants in BPD group were further assigned into mild, moderate and severe BPD groups. Plasma NTproBNP were measured on 14 d, 21 d, 28 d, 35 d, 42 d and 49 d after birth. Repeated-measures ANOVA was used to determine the differences of NTproBNP at different time points in each group.Results:A total of 190 infants were enrolled, including 36 cases in BPD group (18, 13 and 5 cases in mild, moderate and severe BPD group, respectively) and 154 cases in non-BPD group. The gestational age, birth weight and 5-min Apgar score in BPD group were lower than non-BPD group. BPD group had significantly higher incidences of retinopathy of prematurity, patent ductus arteriosus and necrotizing enterocolitis and significantly longer duration of invasive mechanical ventilation and noninvasive ventilation than non-BPD group ( P<0.05).No significant differences existed in NTproBNP levels between BPD group and non-BPD group on 42 d and 49 d ( P>0.05). At other time points, NTproBNP levels in BPD group were significantly higher than non-BPD group ( P<0.05). NTproBNP level in severe BPD group was the highest on 14 d. No significant differences existed in NTproBNP levels between mild and moderate groups on 28 d ( P>0.05). At other time points, NTproBNP in severe BPD group was higher than mild and moderate BPD groups ( P<0.001). The receiver operating characteristic curve analysis showed the best cut-off value of NTproBNP was 982 pg/ml on 14 d (AUC=0.907, 95% CI 0.831~0.983). Conclusions:VLBWI/ELBWI with BPD have higher levels of NTproBNP. And the more severe of BPD, the higher the NTproBNP level. NTproBNP has certain predictive values for BPD in VLBWI/ELBWI.

Multi-centre clinical trials on PET/CT brain imaging are complex to organize and require careful co-ordination and management. This article describes considerations, which are necessary when designing and starting a multi-centre clinical trial on PET/CT brain imaging, based on guidelines and multi-center clinical brain imaging studies, providing references for further studies.