This study aims to investigate whether the nucleoprotein (NP) of severe fever with thrombocytopenia syndrome virus (SFTSV) can impact the cellular immunity of host cells. Gene segments that encode the NP and non-structural protein (NSs) of SFTSV were inserted into eukaryotic expression vector VR1012. Host proteins that interact with NP and affect immunity were identified with co-immunoprecipitation (IP), SDS-PAGE, mass spectrometry (MS), and Western blot. Co-localization of NP and the identified host proteins was confirmed by confocal microscopy. A 60kD SSA/Ro, a protein related to immunity, interacted with NP, as found by IP and MS. Confocal microscopy showed that NP and SSA/Ro were co-localized in cytoplasm. These results indicated that SFTSV NP may specifically bind to 60kD SSA/Ro and cause a series of immune responses and clinical symptoms.
Bunyaviridae Infections ; genetics ; metabolism ; virology ; HEK293 Cells ; Humans ; Nucleoproteins ; genetics ; metabolism ; Phlebovirus ; genetics ; metabolism ; Protein Binding ; Ribonucleoproteins ; genetics ; metabolism ; Viral Proteins ; genetics ; metabolism

To evaluate the adjuvant effect of recombinant enterovirus 71 (EV71) subunit vaccine formulated with chitosan, rabbits were orally immunized with recombinant VP1 (rVP1) or rVP1 mixed with chitosan adjuvant. Levels of virus-specific IgG and IgA antibodies in sera, mucosal wash buffer (intestine, nasal cavity, and lung), and feces were determined by indirect enzyme-linked immunosorbent assay (ELISA). The titers of neutralizing antibodies against EV71 were determined using cytopathic effect-based neutralizing assay, and levels of cytokines (IFN-gamma and IL-4) secreted from in vitro-cultured rabbit splenic lymphocytes under antigen stimulation were also determined by ELISA. Results showed that immunization with rVP1 alone could only induce low levels of serum IgG and mucosal IgA, while rVP1 combined with chitosan adjuvant were able to induce significantly higher levels of antibodies, rVP1 can only induce neutralizing antibodies when used in combination with chitosan. Levels of IFN-gamma and IL-4 in the group immunized with rVP1 plus chitosan were significantly higher than those in the group immunized with rVP1 only or those in the control groups. Our study lays the foundation for development of oral VP1 vaccine against EV71 infection.
Adjuvants, Immunologic ; administration & dosage ; Animals ; Antibodies, Viral ; immunology ; Chitosan ; administration & dosage ; immunology ; Enterovirus A, Human ; genetics ; immunology ; Enterovirus Infections ; immunology ; prevention & control ; virology ; Female ; Humans ; Rabbits ; Vaccination ; Vaccines, Subunit ; administration & dosage ; genetics ; immunology ; Viral Proteins ; administration & dosage ; genetics ; immunology ; Viral Vaccines ; administration & dosage ; genetics ; immunology

For diabetes mellitus, little research has been done on the tissue-based or cell-based drug screening model, which has advantages over traditional animal diabetic model in high specificity, high screening volume, low cost and simple manipulation. Considering that the maintenance of complete islet tissue structure is the prerequisite for islet cells to perform their functions normally, an in vitro islet-based drug screening model for diabetes mellitus was established and evaluated. Pancreatic islets were isolated from 3 weeks old mice of either sex by collagenase digestion and density gradient centrifugation as prescribed by Ramanadham S. The volume of 0.1% (W/V) collagenase IV, 0.1% (W/V) Hyaluroridase and 0.1% (W/V) DNase I were 4 times, 2 times and 1 times that of the islets to be digested. And a 2 hours' cold digestion at 4 degrees C was followed by a 10 minutes' warm digestion at 37 degrees C. Under the optimized digestion condition, the islet recovery could be increased by 10%. The isolated islets could survive 6 weeks in vitro and show stable insulin secretion in the first 10 days after inoculation. The obtained islets were cultured in RPMI-1640 medium at 37 degrees C with 5% CO2. Then a diabetic model was established by selecting streptozotocin (STZ) as the evocator and nitric oxide (NO) as the responding index. After 1 day's inoculation, islets culture was treated with STZ, whose concentration ranged from 0 to 5.0 mmol/L. NO was measured by a colorimetric assay at 540nm based on the Griess reaction for 10 min with 0.1 mL Griess reagent and 0.1 mL culture supernatants. Insulin secretion was assayed by RIA methods. Due to the islets-related inoculation variations, NO release and insulin content were both expressed as a percentage of the value recorded in basal experiment which was in the only presence of Krebs culture medium. It was testified that the amount of NO released from islet itself remained steady at 30-35 mmol/L regardless of the changes of STZ concentration from 0 to 5.0 mmol/L. However the NO content in the supernatants of islets culture had close relationship with STZ concentration. This indicated that in this STZ-induced islet diabetic model, NO mainly comes from STZ when it dissolves in water. On the other hand, when STZ changed from 0 to 5.0 mmol/L, the dose-dependent relationship between NO content and insulin secretion showed that the increase of NO came along with the decrease of insulin secretion, which is an important symbol of islet function. As a kind of oxidative free radical, NO is capable of impair islet cells. Thus, NO is a reliable responding index of the model. The optimal STZ concentration in the model is finally determined to be 5.0 mmol/L, under which condition the NO content and insulin secretion is 10.81 times and 0.43 times that in the medium before STZ is added. So if anything is effective in lowering the NO content in the culture, it could protect islets cells from the oxidative attacks of NO. Finally, as an application of the model, the scavenging effect of KOSCr on NO was studied. In a series of KOSCr with different chromium content, all had shown better NO scavenging effects than KOS itself, which could give us an enlightenment of the influence of chromium ion on oligosaccharide. And 1 g/mL KOSCr with 3.519% chromium content can significantly inhibit the NO formation. This has lain a theoretic basis for the research of KOSCr bioactivity and quality control. These results suggested that the STZ-induced diabetic islet model which is impaired by NO free radical can be used effectively, fast and conveniently when screening potential diabetes drugs.
Animals ; Chromium ; pharmacology ; Diabetes Mellitus, Experimental ; metabolism ; Disease Models, Animal ; Female ; In Vitro Techniques ; Insulin ; metabolism ; Islets of Langerhans ; drug effects ; metabolism ; Male ; Mice ; Mice, Inbred ICR ; Nitric Oxide ; metabolism ; Streptozocin ; pharmacology

BACKGROUNDMagnetic resonance spectroscopy (MRS) is one method that can examine noninvasively the alive specimen of the organ, metabolism of the organ and cell, and the biochemistry change. MRS provides the biochemistry information that may be used to diagnose tumors or differentiate the malignant tumor from benign. The objective of this study is to investigate the benign and malignant bone and soft tissue tumors by 1H-MR spectroscopy ((1)H-MRS) on a 3 Tesla MR scanner, then to assess the usefulness of (1)H-MRS in diagnosing bone and soft tissue tumors and distinguishing benign from malignant tumors.

METHODSFifty-six patients with bone and soft tissue tumors proved clinically and pathologically were examined with (1)H-MRS. (1)H-MRS was performed to study malignant musculoskeletal tumors, benign tumors and normal muscle adjacent to lesions to analyze the characteristics, and single-voxel point-resolved spectroscopy sequence was used. Proton brain exam-single voxel of (1)H-MRS which directly appeared in the spectrum, was observed to find the peak height of choline compounds (Cho) opposite to the creatine (Cr), and whether there was a Cho peak. Metabolite values were calculated automatically from the area under each metabolite peak by the Functool 3.1 software. Metabolite ratios of Cho/Cr were manually calculated. Then according to the results, it was judged whether there existed benign or malignant tumors. The Kappa statistical test was used to analyze the MRS results, the histopathology data and the surgical situation. Statistics processing was performed using the software package SPSS11.5 for Windows.

RESULTS(1)H-MRS spectra style of bone and soft tissue tumors was different from that of normal muscle, and differences also existed between benign and malignant tumors. Choline level in malignant tumor was markedly higher than that in benign tumors. Cho/Cr in malignant tumor was higher than in benign tumor significantly (P < 0.05). The true positive rate of bone and soft tissue between benign and malignant tumors was 34/36, the true negative rate was 15/18, the false positive rate was 3/18 and the false negative rate was 2/36. Therefore in the group, sensitivity of the (1)H-MRS was 94% (34/36), specificity was 83% (15/18), positive predictive value was 92% (34/37), negative predictive value was 88% (15/17) and the accuracy rate was 91% (49/54). The MRS results and the histopathology inspection conclusions had very good uniformity. The kappa value was 0.76 +/- 0.10 (P < 0.01).

CONCLUSIONSThe increase of Cho level measured by (1)H-MRS is related to the bone and soft tissue malignant tumor. Cho/Cr in malignant tumor was higher than in benign tumor, so they will play a vital role in the diagnosis and differential diagnosis of bone and soft tissue tumors.

Adolescent ; Adult ; Aged ; Brain Neoplasms ; diagnosis ; Choline ; metabolism ; Creatine ; metabolism ; Female ; Humans ; Magnetic Resonance Imaging ; Magnetic Resonance Spectroscopy ; methods ; Male ; Middle Aged ; Soft Tissue Neoplasms ; diagnosis ; Young Adult

OBJECTIVETo determine the effects of berberine hydrochloride on the expressions of interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) in periodontal tissues in rat periodontitis model.

METHODSBased on the successful rat periodontitis model, the experimental rats were randomized into different groups followed by oral treatment with berberine hydrochloride for 1, 2, 3, 4 weeks and then the rats were sacrificed and analyzed. Pathological assay and HE staining were used to detect the general conditions and pathological changes of rat periodontal tissues. And immunohistochemical staining was conducted to determine the expressions of IL-1beta and TNF-alpha in rats periodontitis model periodontal tissues.

RESULTSThe levels of IL-1beta and TNF-alpha in the periodontitis tissues were significantly higher than that in the control group. Treatment with berberine hydrochloride decreased the levels of IL-1beta and TNF-alpha in periodontitis tissues (P<0.05). Moreover, the general conditions and pathological changes in the control group and groups treated with berberine hydrochloride were much better than that in periodontitis groups.

CONCLUSIONBerberine hydrochloride inhibited the expression of IL-1beta and TNF-alpha in periodontal tissues in rats periodontitis model and promoted the regeneration of the periodontal tissues. This study suggested that berberine hydrochloride may have potential clinical application.

Animals ; Berberine ; Interleukin-1beta ; Male ; Periodontitis ; Rats ; Tumor Necrosis Factor-alpha

OBJECTIVETo explore the angiogenesis and its maturation of hepatocellular carcinoma (HCC) and its correlation with deoxyhemoglobin parameters R2 * and T2 * values and the lesion/muscle R2*, T2 * ratio by using noninvasive magnetic resonance imaging (MRI).

METHODST2 *, R2 * values and the lesion/muscle R2 *, T2 * ratio in tumor periphery and center were calculated via series T2 * images in a total of 31 patients with surgically and pathologically confirmed HCC. After surgery, all sections were obtained from the specimen periphery in accordance with the MR analyzed areas. Continuous slices of each lesion were stained with hematoxylin-eosin (HE), and immunohistochemical staining was performed in vascular endothelial growth factor (VEGF), Flk-1, proliferating cell nuclear antigen (PCNA), CD34, and alpha smooth muscle actin (SMA). The expressions of VEGF, Flk-1, and PCNA index (PI) were evaluated. According to CD34 and SMA, some vascular parameters, including number, mean vessel area, total vessel area, circumference, diameter, distance between adjacent vessels, and variety index of microvessel and mature vessel, were calculated with a computed analysis system. The amounts of arterioles and veinlets, mature vessel index, and mean perfused fraction (mPF) were also recorded. All vessel parameters were compared with the calculated values of MRI.

RESULTSR2 * value or lesion/muscle R2 * ratio decreased and T2 * value or the lesion/muscle T2 * ratio increased in HCC when compared with hepatic parenchyma (P < 0.05); however, those values between lesion periphery and center and among different pathological grades were not significantly different (P > 0.05). T2 * value and the lesion/muscle T2 * ratio significantly decreased when the expression of VEGF was positive (P < 0.05). T2 * value was negatively correlated with microvessel amount (P = 0.047, r = - 0.639), while T2 * value and the lesion/muscle T2 * ratio were positively correlated with mPF (P = 0.040, r = 0.655; P = 0.048, r = 0.40, respectively). R2 * value was also positively correlated with mean area (P = 0.028, r = 0.688), total area (P = 0.021, r = 0.712) or circumference (P = 0.037, r = 0.663) of microvessel, and negatively correlated with mPF (P = 0.024, r = - 0.702). Meanwhile, the lesion/muscle R2 * ratio was positively correlated with mean area (P = 0.043, r = 0.647) and circumference (P = 0.026, r = 0.694) of microvessels.

CONCLUSIONR2 * or T2 * value may be influenced by the variation of deoxyhemoglobin caused by the heterogeneity of angiogenesis.

Adult ; Aged ; Carcinoma, Hepatocellular ; blood supply ; Female ; Hemoglobins ; Humans ; Liver ; pathology ; Liver Neoplasms ; blood supply ; Magnetic Resonance Imaging ; methods ; Male ; Middle Aged ; Neovascularization, Pathologic

Objective To study the correlation between fractional anisotropy(FA)and tumor microarchitecture(MVD,VEGF and celluarity).Methods Fouteen gliomas(5 grade Ⅰ and Ⅱ,4 grade Ⅲ, 5 grade Ⅳ)confirmed histo-pathologically were performed on diffusion tensor imaging(DTI)using a GE Signa Excite Ⅱ 3.0 T MR scanner(8-channel head coil,SE echo planner imaging(EPI),thickness:5 mm, spacing:0,directions:25,B values:0 and 1000 s/mm~2,TR 6000 ms,TE minimum,FOV:240 mm? 240 mm,image matrix 128?128,NEX 2).Postprocessing was done using a DTI specific software to gain FA image.ROIs were drqwn in tumor parenchyma and the value of FA was recorded.The positive expression of VEGF and CD34 was shown using immuno-histochemistry method.The VEGF,MVD,and cellularity of every slices were recorded.Pearson correlation analysis was used.Results FA(which is 0.102?0.080 in grade Ⅰ and Ⅱ,0.171?0.037 in grade Ⅲ,0.200?0.021 in grade Ⅳ)has the trend to raise with the increasing grade of astrocytomas.FA has significant positive correlation to MVD(40/HP in grade Ⅰ and Ⅱ, 86/HP in grade Ⅲ,101/HP in grade Ⅳ),VEGF(8% in grade Ⅰ and Ⅱ,47% in grade Ⅲ,55% in grade Ⅳ),and cellularity(104/HP in grade Ⅰ and Ⅱ,160/HP in grade Ⅲ,265/HP in grade Ⅳ).The correlation coefficients between FA and VEGF,MVD,and cellularity were 0.748,0.668,0.625 respectively.Conclusion As a new imaging method,DTI can reveal the microarchitecture in gliomas and be value of distinguishing gliomas of different grade.DTI provides a new method of precise diagnosis to glioma preoperatively.

Objective To evaluate the efficacy and safety of memantine in the treatment of patients with Alzheimer' s disease (AD).Methods This was a 16-week,multi-center,randomized,double blind, placebo-controlled clinical trial (Study 10116).A total of 258 AD patients (MMSE score 5—18) were randomized in a 1:1 ratio into either memantine 10—20 mg/day (MEM,n=128) or placebo (PBO,n= 130) group for 16 weeks.Efficacy was primarily assessed in terms of changes of severe impairment battery (SIB) score in patients from baseline up to SIB assessment in the 16th week (16-week completers set, CS16).While ehanges of MMSE,ADCS-ADL_(19),and NPI (neuropsychiatric inventory) were evaluated as secondary efficacy parameters on both CSI6 and full-analysis set (FAS).Safety was assessed by physical examination,lab assays,ECG,and adverse events.Results 236 subjeets (CS16:MEM n=117,PBO n=119) were eligible for the efficacy assessment.No statistically significant difference between the treatments was observed on the primary and seeondary efficacy analysis,although both treatment groups had a slight increase from baseline in SIB total score.Post hoe evaluation of the data identified two bias factors that had a significant impact on the results of the pre-protoeol specified primary and secondary analyses.In a re-analysis of the data (CS16_(modified),MEM n=94,PBO n=95) excluding patient data affeeted by these factors,memantine-treated patients showed a statistically significant improvement related to placebo in the 16th week on the SIB (MEM 2.2 vs PBO 0.3,P=0.04),MMSE (MEM 1.0 vs PBO 0.1,P=0.03),and ADL (MEM 0.1 vs PBO-1.6,P=0.02) scales,indicating that memantine improved the cognitive function of AD patients and stabilized the activity of daily life.Memantine was well tolerated with an adverse event profile similar to that of placebo.Conclusion This study provides further support for pre-existing data,showing that memantine is efficacious,safe,and well-tolerated in patients with moderate to severe AD.

OBJECTIVETo investigate the correlation between the number and activity of circulating endothelial progenitor cells (EPCs), insulin resistance and severity of coronary lesions in patients with coronary artery disease (CAD).

METHODSPatients with coronary angiography evidenced CAD were divided in insulin resistance group (IR, n = 25) and insulin sensitive group (IS, n = 44) according to insulin level, 25 health volunteers served as control. Circulating EPCs were marked as KDR/CD133+ cells via fluorescence-activated cell sorter analysis. EPCs were also isolated from peripheral blood and cultured in vitro for 7 days, identified by DiI-acLDL uptake and lectin staining methods. EPCs migration activities were determined by modified Boyden chamber assay, EPCs proliferation activities were determined by MTT assay.

RESULTCirculating EPCs number was significantly lower in IR group compared with IS group [(0.34 +/- 0.08) per thousand vs. (0.47 +/- 0.09) per thousand, P < 0.01] and control group (P < 0.05). Both insulin resistance index (r = -0.291, P = 0.01)and Gensini score (r = -0.3984, P = 0.006)were negatively correlated with number of circulating EPCs. Proliferation and migration capacities of EPCs were also significantly lower in IR group compared to those in IS group (all P < 0.05) and control group (all P < 0.05).

CONCLUSIONSInsulin resistance/hyperinsulinemia could aggravate severity of coronary artery lesions via reducing the number and activities of circulating EPCs in patients with CAD.

Aged ; Blood Cell Count ; Cell Adhesion ; Cell Movement ; Cell Proliferation ; Cells, Cultured ; Coronary Angiography ; Coronary Artery Disease ; blood ; pathology ; Endothelial Cells ; cytology ; Female ; Humans ; Insulin Resistance ; Male ; Middle Aged ; Stem Cells ; cytology

UNLABELLEDOBJECTIVE; To evaluate a technique with retained copper needles for the treatment of venous malformations.

METHODSWith 78 venous malformation cases, there were three methods were applied for the treatment respectively, including copper needles in the lesion only, vascular ligation with the copper needles in the lesion, and electrical puncture with the copper needles in the lesion.

RESULTSThere were totally 96% effective rate achieved in this clinical data.

CONCLUSIONSThe retained copper needles technique may be a simple and effective method for the treatment of venous, malformations resulting in vessel denaturation, fibrosis and disappearance of structure.

Adolescent ; Adult ; Child ; Electric Stimulation Therapy ; Embolization, Therapeutic ; instrumentation ; methods ; Female ; Humans ; Male ; Middle Aged ; Needles ; Vascular Malformations ; therapy ; Young Adult