Flammulin, an anti-tumor protein, was purified from the aqueous extract of basidiomes of Flammulina Velutipes. Purified flammulin emulsified with Freund's adjuvant was injected subcutaneously into New Zealand white rabbits. After several immune enhancements, these animals were bled and sera were separated. Antiserum against flammulin in Western blots were applied to determine if flammulin be present in the liquid state culture or fruiting body. The result showed that anti-flammulin serum could recognize the aqueous extract of fruiting body in SDS-PAGE gels under the reducing conditions, no flammulin was detected in mycelia of Flammulina Velutipes.

Objective To determine the effect of fibroblast growth factor (FGF) on endothelia cell (EC) proliferation in vitro,TNP-470 and dexamethasone (Dex) and explore the possible mechanisms of FGF stimulating EC proliferation. Methods Cultured human umbilical vein endothelial cell (HUVEC) was divided into two groups: control group (Group DMEM) and experimental groups (Group FGF,Group Dex,Group TNP-470,Group FGF+Dex,Group FGF+TNP-470). The EC proliferation was quantified by a colorimetric assay using MTT reagent,the relative cell numbers at every time point of EC cycle analyzed with flow cytometer,and the expression of nuclear factor-kappa B p65 (NF-?B p65) and ki-67 detected with immunohistochemical method of SABC. Results FGF stimulated significantly EC proliferation,raised proliferation index (PI) and enhanced the expressions of NF-?B p65 and ki-67 in nuclear. TNP-470 reduced PI and TNP-470 and Dex suppressed EC proliferation and finally decreased the expressions of NF-?B p65 and ki-67 in nuclear. FGF could facilitate the inhibitory effect of TNP-470 and Dex. Conclusions The possible mechanism of EC proliferation stimulated by FGF is that FGF can activate NF-?B to promote the synthesis of DNA and EC mitosis. TNP-470 and Dex suppress proliferation and differentiation of EC but FGF may reverse such suppressive effect.

Theultraperformanceliquidchromatographycoupledwithmassspectrometry(UPLC-MS)was used to investigate the metabolic difference of the decoction of Radix Aconiti Preparata ( RAP ) and its co-decoctions with Radix Paeoniae Alba ( RAP-RPA ) or Radix Stephaniae Tetrandrae ( RAP-RST ) in rat intestinal bacteria. The principal component analysis ( PCA) of the relative contents of Aconitum alkaloids after metabolism was performed by SIMCA-P software. The score plots of PCA could successfully distinguish the three groups of RAP, RAP-RPA and RAP-RST. The result indicated that the differences of biotransformation among the groups of PAP, RAP-RPA and RAP-RST were significant. With the loading plot and independent-samples T test, seven relevant markers with the significant differences were found in the group of RAP-RPA, six relevant markers were obtained in the group of RAP-RST. The relative content of four markers in RAP-RPA was higher than that in RAP, and one marker in RAP-RST was higher than that in RAP. The relative contents of other markers were all lower than that in RAP. These markers may be the effective substance for explaining the different effects of Radix Aconiti Preparata before and after combination with Radix Paeoniae Alba and Radix Stephaniae Tetrandrae.

Objective:To characterize the biological activity of fibroblasts on the surface of titanium alloy sheets with different ridge widths by investigating the effects of ridge widths on the adhesion, proliferation and differentiation of fibroblasts.Methods:Five groups of titanium sheets with ridge widths of 50 μm, 80 μm, 100 μm, 150 μm and 200 μm were prepared, with all the groove depths being 10 μm. The titanium sheets with no ridges were taken as a control group. After fibroblasts were incubated on the sheets, states of their adhesion were observed by scanning electron microscopy (SEM) at different time points. CCK-8 cell proliferation test and immunofluorescence staining were used to observe proliferation and shape of the cells. The effects of ridge widths on adhesion of fibroblasts were evaluated by Vinculin immunofluorescence staining, and the effects of ridge widths on expression of α-smooth muscle actin ( α-SMA) by immunofluorescence. Results:SEM showed that the cells adhered to the ridges on the titanium sheets 48 hours after inoculation. In the groups with smaller ridge widths (from 50 μm to 150 μm), the cells were slender in shape and grew along the ridge direction. CCK-8 indicated that different ridge widths had no significant effect on the proliferation of fibroblasts between the 6 groups ( P>0.05). Immunofluorescence staining showed that the cells arranged in an orderly direction along the ridges; the long axis of the cells in the 50 μm group showed the best consistency with the extending direction of the ridge, with significant differences among the 6 groups ( P<0.05). The Vinculin test found that the secretion of cell adhesion protein was concentrated in the ridge and semi-quantitative analysis showed that the 50 μm group had the most Vinculin secretion, with significant differences among the 6 groups ( P<0.05). The α-SMA test showed that the ridge width had a regulatory effect on the myogenic differentiation of fibroblasts, and the 50 μm group had the strongest expression of α-SMA, with significant differences among the 6 groups ( P<0.05). Conclusions:Modification of ridges on the surface of titanium sheets may affect arrangement, adhesion and myogenic differentiation of fibroblasts. The ridges of 50 μm in width may lead to stronger polarized arrangement of fibroblasts, more secretion of adhesion-related protein and more pronounced myogenic differentiation of fibroblasts.

OBJECTIVETo explore the role of S100B protein in the early diagnosis, treatment, and prognosis judgement of craniocerebral injury.

METHODSIn this study, we reviewed the domestic and foreign research reports about the relationship between S100B protein and craniocerebral injury.

RESULTSThe concentration of S100B protein had a different increase based on the degree of injury in early stage after craniocerebral injury, and the increasing degree of S100B protein showed a positive correlation with the grading of pathogenetic condition and prognosis of craniocerebral injury.

CONCLUSIONSS100B protein may be taken as a specific index of early diagnosis, grading of pathogenetic condition, and prognosis judgement after craniocerebral injury. To grasp and regulate the mechanism of neurotoxicity and to elucidate the therapeutic effect of S100B protein will be a research direction in clinical treatment of craniocerebral injury.

Craniocerebral Trauma ; cerebrospinal fluid ; diagnosis ; Humans ; Nerve Growth Factors ; cerebrospinal fluid ; Prognosis ; S100 Calcium Binding Protein beta Subunit ; S100 Proteins ; cerebrospinal fluid

Adult ; Humans ; Knee Injuries ; surgery ; Male ; Menisci, Tibial ; surgery ; Transplantation, Homologous ; methods

OBJECTIVETo develop a therapeutic vaccine against human tumors associated with human papillomavirus type 16E6E7 (HPV16E6E7) which is modified from a Chinese patient of the cervical cancer which possessing the antigenicity and no transforming activity, and explore more active vaccine for inducing cellular immunity with mouse co-stimulatory molecular B7-1 gene.

METHODSThe modified E6E7 gene expression plasmid pVR1012-fmE6E7 was constructed and transfected Cos-7 cells, and the E7 protein specific expression was testified by immunofluorescence assay. C57BL/6 mice were immunized intramuscularly with pVR1012-fmE6E7 alone or in combination with B7-1 gene expression plasmid (pcDNA3.1-B7-1). The activity of cytotoxic T lymphocytes (CTLs) was analyzed with 51Cr specific release assay and the specific antibody in sera was analyzed by indirect ELISA. HPV16 positive C57BL/6 tumor cells C3 were inoculated subcutaneously in the vaccinated mice to assay the growth of transplanted tumors.

RESULTSThe specific CTLs and antibody from immunized mice were induced efficaciously by the E6E7 gene immunization, and co-administration of B7-1 gene could significantly enhanced the CTLs immune responses of fmE6E7, and protected 33% immunized mice against C3 tumor cells challenge. In contrast, all the mice immunized only with fmE6E7 gene developed transplanted tumors after C3 cells challenge. There was no difference in E7 specific antibody responses between mice immunized with the E6E7 gene only and co-administration with B7-1 gene.

CONCLUSIONSThe modified E6E7 gene can be used as target gene for developing DNA vaccine, and B7-1 gene may represent an attractive adjuvant for enhancement of the specific cellular immune responses.

Animals ; Antibodies, Neoplasm ; immunology ; B7-1 Antigen ; genetics ; immunology ; Female ; Mice ; Mice, Inbred C57BL ; Neoplasm Transplantation ; Oncogene Proteins, Viral ; genetics ; immunology ; Papillomavirus E7 Proteins ; Repressor Proteins ; T-Lymphocytes, Cytotoxic ; immunology ; Transfection ; Uterine Cervical Neoplasms ; immunology ; pathology ; virology ; Vaccines, DNA ; immunology

Objective To analyze the molecular epidemiological characteristics and genotype distribution of norovirus in Zhenjiang region during 2015 and 2016.Methods The fecal samples from diarrhea patients in Zhenjiang region were collected,and the expression level of norovirus capsid protein gene was detected by reverse transcription PCR (RT-PCR).The PCR products were validated by gene sequencing.Then,phylogenetic tree was constructed with Mega software,and sequence evolution and genotype were analyzed.Results The positive rate of norovirus infection was 5.30％ (85/1 605).Among them,the detection rates of G Ⅰ genogroup and G Ⅱ genogroup were 0.87％ (14/1 605) and 4.55％ (73/1 605),respectively,and 2 samples were detected both of G Ⅰ and G Ⅱ genogroups.The highest positive rate of G Ⅱ genogroup occurred in adolescent patients aged from 10 to 20 years (17.80％).During 2015,the detection rate of norovirus infection was the highest in January,and then in November and February.During 2016,the detection rate of norovirus infection was the highest in December,and then in March and April.The main genotypes of norovirus in 2015 and 2016 were new G Ⅱ.17 variant (63.89％) and G Ⅱ.4 Sydney_2012 strain (35 ％),respectively.Conclusion The infection of norovirus in Zhenjiang region appears G Ⅱ as main genogroup,adolescent as main patients,November to April of the next year as main epidemic seasons,and G Ⅱ.17 variant and G Ⅱ.4 Sydney_2012 strain as main epidemic genotypes.

Urine samples from 72 patients with diabetic nephropathy (DN) were analyzed and compared to those from 33 diabetic patients without albuminuria and 29 normal controls,using SELDI-TOF-MS (surface enhanced laser desorptiort/ionization time-of-flight mass spectrometry) and Biomarker Patterns Software,to identify differences in protein profile,generate a tree analysis pattern and evaluate the validity of the decision tree.The intensities of 6 peaks detected appeared upregulated,while 11 peaks downregulated,in DN group as compared to nonDN groups more than 2 folds (P

OBJECTIVETo evaluate the advantages, disadvantages and their indications of total nasal reconstruction with different techniques.

METHODSA series of total nasal reconstruction were treated with four methods from 1975 to 2003. These methods were tubed flap of arm,midline forehead flap with skin graft, midline forehead flap with bilateral frontotemporal flaps for repairing the donor site, and expanded forehead flap.

RESULTSAll of the patients were treated successfully. The shape and function of the reconstructed noses were satisfactory. However, the traditional forehead flap with skin graft may leave a unsightly big and black scar on the forehead. The technique of the tubed flap of arm could provide enough tissue without remaining forehead scar and be easily shaped, but it required long period, multiple procedures and body fixation for three weeks.

CONCLUSIONSMidline forehead flap with bilateral frontotemporal flaps for repairing the donor site may be good for small nose reconstruction while expanded forehead flap could reconstruct a big nose. Tubed flap of arm may be used to the patients who do not wish to leave any scar on the forehead. Forehead flap with skin graft to repair the donor sit- should generally be avoided for nose reconstruction.

Adolescent ; Adult ; Female ; Humans ; Male ; Middle Aged ; Nose ; surgery ; transplantation ; Nose Deformities, Acquired ; surgery ; Reconstructive Surgical Procedures ; methods ; Reproducibility of Results ; Retrospective Studies ; Skin Transplantation ; methods ; Surgical Flaps ; Treatment Outcome