Objective To measure the expression of granulocyte colony-stimulating factor receptor (G-CSFR) in human melanocytes and to evaluate the biologic effect of recombinant human granulocyte colony-stimulating factor (rhG-CSF) on human melanocytes.Methods Melanocytes were obtained from circumcision specimens of healthy males,and neutrophils were isolated from heparin-andcoagulated peripheral blood of healthy human followed by a primary culture.Then,the melanocytes in third passage were cultured with or without the presence of various concentrations (200,400,600,800 μg/L) of rhG-CSF for 72 hours.The growth and morphology of melanocytes were observed.Flow cytometry was performed to detect the expression of G-CSFR in untreated human melanocytes,neutrophils and erythroleukemia cells (HEL 92.1.7).Western blot and reverse transcription PCR (RT-PCR) were carried out to measure the expression of G-CSFR protein and mRNA respectively in the neutrophils,HEL 92.1.7 cells,treated or untreated human melanocytes.Methyl thiazolyl tetrazolium (MTT) assay was performed to evaluate the proliferation,and dopa-oxidation assay to estimate the tyrosinase activity,of treated melanocytes.Results The expression rate of G-CSFR was 76.81％ ± 10.70％ in human melanocytes,significantly higher than that in the HEL 92.1.7 cells (2.53％ ± 1.54％,P ＜ 0.01 ),but lower than that in the neutrophils (85.76％ ± 15.71％,P ＜ 0.05).Both G-CSFR protein and mRNA were expressed in melanocytes,and there was no significant differences in the expression level of G-CSFR protein and mRNA among melanocytes treated with different concentrations of rhG-CSF (both P ＞ 0.05).The expression levels of G-CSFR protein and mRNA in the melanecytes were significantly higher than those in the HEL 92.1.7 cells (both P ＜ 0.01 ),but lower than those in the neutrophils (P ＜ 0.05 or ＜ 0.01 ).rhG-CSF at 200-800 μg/L displayed a significantly promotive effect on the proliferation of melanocytes (P ＜ 0.01 or ＜ 0.05 ),and the effect was in a dose-dependent manner when rhG-CSF ranged from 200 to 600 μg/L (P ＜ 0.01 ).The rhG-CSF at 600 μg/L and 12-O-tetradecanoyl-phorbol-13-acetate (TPA) at 20 μg/L showed an equivalent effect on the proliferation of melanocytes (164.04％ ± 13.0％ vs.165.62％ ± 10.6％,P ＞ 0.05).However,rhG-CSF from 200 to 800 μg/L had no significant impact on the tyrosinase activity of melanocytes (all P ＞ 0.05 ).Conclusions G-CSFR is expressed in human melanocytes. rhG-CSF can promote the proliferation of cultured human melanocytes,but has no obvious influence on the tyrosinase activity of melanocytes.

Objective:To investigate whether platelet and coagulation-related indicators can be used as early indicators of severity of acute pancreatitis.Methods:A total of 142 patients with acute pancreatitis admitted to Beijing Friendship Hospital from 2017 to 2018 were included in this study. According to the Ranson score, they were divided into mild group and severe group. Severe pancreatitis was used as the outcome index. Univariate logistic regression analysis and receiver operating characteristic (ROC) curve analysis were performed on single indicators with statistically significant differences between mild and severe patients to determine the discriminative value of disease severity.Results:According to the Ranson score, 142 patients were divided into severe and mild groups, 43 patients with severe disease, and 99 patients with mild disease. There was a statistically significant difference in platelet, fibrinogen (FIB), and D-Dimer between the two groups on the first day of admission ( P<0.05). The area under ROC curve and 95% CI of platelets (PLT) , FIB and D-Dimer were 0.61 (0.52, 0.71), 0.70 (0.59, 0.80) and 0.72 (0.62, 0.82), respectively. Multivariate logistic regression models of PLT, FIB and D-Dimer were fitted, and ROC curve was analyzed. The area of ROC curve of PLT combined with D-Dimer was 0.74 (0.65, 0.83), and the area of ROC curve of FIB combined with D-Dimer was 0.75 (0.66, 0.85). Conclusions:Platelet, FIB and D-Dimer can be used as independent risk factors to judge the severity of pancreatitis. The early predicative value of FIB combined with D-Dimer on the severity of the disease is higher than that of the PLT combined with D-Dimer.

Human bocavirus (HBoV) 1-4 have been detected both in respiratory and stool samples since the first HBoV was discovered in 2005. HBoV-1 is mostly associated with respiratory infection, while HBoV 2-4 are usually associated with intestinal tract infection. A variety of signs and symptoms have been described in patients with HBoV infection, including cough, wheezing, pneumonia, and diarrhea, but the research on pathogenic mechanism of HBoV is limited because HBoV cannot be cultured in vitro due to the lack of appropriate host cells. Three-dimensional epithelial cell culture, reverse genetics, and viral metagenomics are identified as novel tools that may promote the research on pathogenic mechanism of HBoV and the discovery of new viruses. This review summaries currently available diagnostic approaches such as electron microscopy, cell culture, PCR, and immunoassay in order to provide a method reference for indepth research on HBoV.
Animals ; Human bocavirus ; genetics ; growth & development ; isolation & purification ; pathogenicity ; Humans ; Parvoviridae Infections ; diagnosis ; virology ; Viral Proteins ; genetics ; metabolism ; Virology ; methods ; Virulence ; Virus Cultivation

Objective:To investigate the in vitro effects of platelet-rich plasma(PRP) on human periodontal ligament fibroblasts(PDLFs). Methods: Various concentrations of PRP (10, 50, 100, 200, 300, 500 ml/L) were applied to primary cultures of human PDLFs. MTT assays were utilized to assess cell proliferation ability. Migration was determined by assessing the cell response to a concentration gradient with Transwell chamber. Differentiation was assessed using alkaline phosphatase (ALP) kit. Results: A beneficial effect on proliferation was observed, especially in response to 200 ml/L PRP.PRP had stimulatory effects on the migration of human PDLFs. PRP facilitated differentiation of PDLFs. Conclusion: PRP can exert a positive effect on human PDLFs,but this effect is concentration specific, while higher concentrations is not necessary to result in optimal outcomes.

Objective To investigate the effect of extracorporeal cjrcuit on the concentration of sufentanil the priming solution.Methods The extracorporeal circuit (ECC) of Xi-jing-90 type (group A) was used in the study,while incontrol group (group B) a glass container was used.The ECC and glass container were filled with priming solution (succinylated Gelatin 1000 ml + lactated Ringer's solution 1000 ml).Sufentanil 15 μg was then added to the priming solution (the final concentration was 7.5 ng/ml).The priming solution was circulated in the closed ECC or stirred in the glass container.The concentration of sufentanil in the priming solution was determined at 3,5,10,20,30,40,50,60,70,80 and 90 min after addition of sufentanil by gas chromatography.Results The sufentanil concentration in.the priming solution decreased in group A at the different time points respectively as compared with group B (P ＜ 0.05).Conclusion Sufentanil can significantly be absorbed by the extracorporeal circuit.

Objective To investigate the feasibility and promising applications of percutaneous transhepatic lymphosonography in detecting sentinel lymph nodes(SLNs).Methods Twenty five rabbits with VX2 tumor were included in this study.0.05 ml SonoVue was injected into the liver parenchyma at 12,3,6,9 points around the VX2 tumor.The situation of contrast-enhanced lymph-vessel emited from injected point and lymph nodes in hepatic portal or around tumor was observed,and then the position of the lymph nodes were detected with the help of the mark on the surface of the portal vein,caput pancreatis,collum vesicae biliaris.Methylene blue (MB) was injected in the same way as above.The injected points were massaged for five minutes,and then executed the experimental rabbits.The lymph nodes enhanced and all the lymph node dyed or not were taked out for recorded and pathologic examination.Results 34 SLNs were conformed by operation and pathological diagnosis in all the rabbits.All SLNs were confirmed pathologically,28 lymph nodes which were checked out by percutaneous transhepatic lymphosonography were all SLNs.In all the 31 lymph nodes which were checked out by MB,25 lymph nodes were SLNs and the rest were the second degree lymph nodes.The detection rate of percutaneous transhepatic lymphosonography (82.4％) and the MB (91.2％) showed no significant difference(P =0.169).There were 6 SLNs enhanced uniformitily in which 2 SLNs encroach by cancer cell and 22 enhanced asymmetrically in which 21 SLNs encroach by cancer cell.The sensitivity,specificity and accuracy of percutaneous transhepatic lymphosonography to detcect the SLNs benign or malignancy was 95.5％ (21/28),66.7％(4/6) and 89.3 ％ (25/28).Conclusions Percutaneous transhepatic lymphosonography is a reliable and noninvasive method to detect and estimate the SLNs of hepatic cancer.

Objective To explore the effects of modified tubo-uterine implantations performed on women with proximal tubal occlusive infertility after femal sterilization with mucilago phenol.Methods Two hundred and eight infertile women who were admitted to the Second Affiliated Hospital of Sun Yat-sen University between 1986 and 2004 were included.They all accepted modified tubo-uterine implantation after occlusion of fallopian tubes with mucilago phenol.Results It was found that the occlusions were all located in the interstitial portion or isthmic portion of the fallopian tubes.Different degrees of pelvic adhesions were found in 65 cases.Fifty-seven cases were slightly adhesive,seven cases were of moderate degree and one case was severe.One hundred and ninety-nine cases were followed up after operations(95.7%).One hundred and ninety-three women accepted hydrotubation in the following month just after the operation and 185 women were found to be unobstructed(95.8%).One hundred and forty-three women became pregnant, the pregnant rate being 71.9%(143/199).One hundred and twenty-five women had term deliveries (87.4%),three women were in early pregnancy and two in midtrimester pregnancy.Eleven women had spontaneous abortion(7.7%).Two women had tubal pregnancy(1.0%).None of the 199 cases had any signs of endometriosis.Conelusions Modified tubo-uterine implantations are quite effective for proximal tubal occlusive infertility.It may be a favorable method for such kind of tubal occlusions.

Protein phosphorylation is one of the most common post-translational modifications (PTMs)in various organisms,which plays critical roles in the regulation of intracellular biological processes,such as cell prolifera-tion,signal transduction,metabolismis and tumorigenesis.However,the low abundance of phosphoprotein in the biological systems poses significant challenges of current analytical techniques.In order to further understand the phosphoproteomics,the roles of phosphorylated proteins in life process,discovery of biomarkers,diagnosis and treatment of disease,enrichment strategies of high efficiency have been developed,including the design of new nanomaterials and combination of a variety of analytical methods,et al.In this paper,we reviewed the develop-ment of enrichment strategies for phosphoproteomics and application of phosphoproteomics in disease.

ObjectiveTo observe the dynamic change of muscle tissue restoration after damaging at different observation time through acoustic radiation force impulse(ARFI).MethodsAccording to different observation time,16 healthy rabbits were randomly divided into four groups,including before injury,1 day after injury,7 day after injury,14 day after injury.A homemade gravity hammer was used to establish damage model of rabbit gastrocnemius,then applied ARFI to observe changes of virtual touch tissue quantification (VTQ) of gastrocnemius.All the rabbits were executed after measurement,then the pathological changes of muscle tissue were observed under microscope.Results The VTQ of damaged muscle group were significantly higher than that of nomal muscle group( P ＜0.01),and VTQ of 1 day,7day,14 day group after injuried had significant difference between two groups (P ＜ 0.01).Through pathological examination,normal musule fibers were continual and there were not swelling and bleeding.One days after injury,muscle fibers were fractured,swollen,bleeding and congestive.7 days after injury,large areas of muscle cells were necrotic,and amounts of calcium salt depositsed.14 days after injury,lots of fiber cells proliferated,and the deposits of calcium obviously reduced.The results of ultrasound elastography were consistent with these of pathological.ConclusionsARFI can direct,noninvasively evaluate the changes of muscle tissue restoration after different time of injury,and provide objective basis for diagnosis and treatment.

Objective To observe the expression of mitochondrial fission protein locus Fis1 and ultrastructural changes in the renal cells of rats with chronic fluorosis,and to reveal the mechanism in mitochondrial damage of the renal cells.Methods Sixty SD rats were randomly divided into 3 groups according to sex and body mass(20 in each group):control group,lower fluoride group and higher fluoride group.All the rats were fed with different doses of sodium fluoride in drinking water(0,10 and 50 mg/L,respectively).Six-month later,the expression of Fisl in renal cells was determined by real-time fluorenscence quantitative PCR and immunohistochemistry technology,the mitochondrial morphology of renal cells was observed under transmission electron microscopy (TEM).Results As compared with the control group(28.70 ± 12.41),Fis1 mRNA levels(91.48 + 34.83 and 582.09 ± 184.69) in renal cells of the lower fluoride and the higher fluoride groups were increased(all P ＜ 0.05).As compared with the control group(10.49 ± 7.66),Fisl protein levels(16.33 ± 10.26 and 21.50 ± 5.24) in renal cells of the lower fluoride and the higher fluoride groups showed a trend of increasing,the higher fluoride group was higher than that of the control group(P ＜ 0.05).By TEM,mitochondrial crest in renal cells of the lower fluoride and the higher fluoride groups was vague or disappeared,mitochondrial division section appeared.Conclusions Fluoride is a kind of toxicant that can cause damage to mitochondrion of renal cells,induce the expression of Fis1 in transcriptional and protein level,and lead to the obstacles of mitochondrial fusion-fission and ultrastructural abnormality of mitochondrion,which may play an important role in mechanism of mitochondrial damage in the renal cells of rats with chronic fluorosis.