Leonurine(SCM-198)was discovered as one of the active constituents of the Herba Leonuri(HL).Now it can be artificially synthesized.Several recent researches has proven that it exhibits anti-inflammatory effect in several systems in animal models and cell culture in vitro.The key mechanism involves downgrading the activity of nuclear transcription factor-κB(NF-κB),thereby inhibiting the phosphorylation of several signal pathways such as PI3K/Akt,MAPK,ERK,and JNK,or upregulating the activity of Nrf2 related pathways,resulting in downregulated expression of inflammatory cytokines such as tumor necrosis factor-α(TNF-α),IL-1β,IL-2,IL-6,IL-8,inducible nitric oxide synthase(iNOS),cyclooxygenase-2(COX-2),chemokines,adhesion molecules,etc.Owing to the advantages of high safety and efficiency,the ease of administration,as well as its effectiveness in many organs and systems,leonurine has a widely prospect for future research and clinical applications.This article reviews the progress in the fundamental research of leonurine in multiple inflammation-related disease,and it could be expect to offer new possibilities for the treatment of these disease.

Objective:To evaluate the effect of low-dose recombinant interleukin-2(rIL-2)therapy on immunocyte subsets and its side effects in children with solid tumor.Methods:A total of 22 children(11 males and 11 females)with solid tumor in our department from December 2012 to November 2017 were selected,with a median age of 9(3-16)years old when starting IL-2 therapy.ALL surgeries and chemotherapy of children had been completed before low-dose rIL-2 therapy,and 17 cases achieved complete remission(CR)and 5 cases achieved partial remission(PR).A low-dose rIL-2 therapy was given 1 month after chemotherapy for 1 year:4 × 105 IU/(m2·d),s.c.for every other day,3 times per week.The immunocyte subsets were detected every 3 months until the end of treatment,meanwhile,disease condition and therapy-related side effects were followed up.Results:After low-dose rIL-2 therapy in 22 children,the absolute values of CD3+T cells,CD3-CD56+natural killer cells,CD3+CD4+helper T cells(Th)and CD3+CD8+cytotoxic T cells were up-regulated remarkably,as well as Th/suppressor T cells(all P＜0.05).While,there were no significant differences in absolute value and proportion of CD4+CD25+CD127-Treg cells during therapy.Among the 17 children who achieved CR before rIL-2 therapy,14 cases continued to maintain CR after therapy,while 3 cases relapsed,and with 2 died after treatment abandonment.The 5 children who achieved PR before low-dose rIL-2 therapy were evaluated CR by PET/CT scan after treatment.In the early stage of low-dose rIL-2 therapy,1 child developed skin rashes at the injection sites,and 2 children ran a slight to mild transient fever.Their symptoms disappeared without any organ damage after symptomatic treatment.Conclusion:Low-dose rIL-2 therapy has good drug tolerance,and changes the distribution of anti-tumor immune-cell subgroup in peripheral blood of children with solid tumor remarkably without up-regulation of absolute value and ratio of Treg cells.

Objective: To investigate the possibility of adipose tissue mesenchymal stem cell-derived microvesicles (ADSC-MVs) to improve the retention volume of fat transplantation. Methods: Human adipose tissue was obtained from 5 healthy female patients aged from 20 to 30 years, who came to the hospital for abdominal liposuction. Adipose tissue mesenchymal stem cells (ADSCs) were acquired by collagenase enzymatic hydrolysis. ADSC-MVs were isolated from the supernatant of cultured ADSCs through ultra-centrifugation, and characterized by transmission electron microscope and PKH26 staining. Sixteen BALB/c-nu nude mice were randomly divided into 2 groups (n=8, mice/group) by random number table method. In EV group, the mice were subcutaneously injected 0.35 ml human fat, with 7 μg ADSC-EVs (final concentration is 20 μg/ml), while 0.35 ml human fat were in blank group. The mice were sacrificed and the grafts were harvested at 1 month and 3 months after transplantation. Weight and volume of transplanted fat were tested. Morphology of adipocytes, fibrosis and necrosis of fat tissue were confirmed by HE staining. Blood vessel density were accessed by CD31 staining.SPSS 13.0 was used for statistical analysis and Student-t test was applied to compare the data from two groups. Results: One month after fat transplantation, the volume of fat grafts in blank group was lower than that in MV group. Besides, lipoliquefaction can be observed in fat grafts of blank group. Fat grafts in MV group had a higher volume and littler necrosis and lipolique faction. Three months after grafting, the volume of fat grafts in blank group [(0.057±0.009) ml] was significantly lower than that in MV group [(0.103±0.015) ml], t=8.117, P<0.001. MV group had an increased number of vessels (29.6±4.0/field) compared with grafts from the Blank group (23.0±2.4/field), t=2.825, P=0.022. Histological analysis revealed that the fat grafts in MV group consisted less fat necrosis and fibrosis than control group. Conclusions ADSC-MVs can improve the retention volume of fat transplantation.

Objective To explore the effect of divided injection of fat grafting and to provide a new sight for the strategy of clinical particle fat transplantation.Methods Adipose tissue was aspirated from the healthy female abdomen by liposuction.In the control group,0.5 ml of adipose tissue was subcutaneously injected into the nude mice.The experimental group was injected with 0.25 ml first,followed by 0.25 ml injection of adipose tissue on the 7th,14th and 30th days.To assess graft retention rate and effectiveness we measured the wet weight and observed the pathological sections with HE or perilipin immunofluorescence.Results The wet weight of the tissue between the experimental group and the control group had no statistical difference,the experimental group had less necrosis and empty tissue than the control group.The proportion of perilipin positive staining tissue in 7 day group had statistical difference from that of the control group.Conclusions The strategy of preinjection of part of the adipose tissue and then supplement of the residual tissue after 7 days may increase the proportion of active adipose tissue in the graft.

Ligusticum chuanxiong is a well-known traditional Chinese medicine plant. The study on its molecular markers development and germplasm resources is very important. In this study, we obtained 24 422 unigenes by assembling transcriptome sequencing reads of L. chuanxiong root. EST-SSR was detected and 4 073 SSR loci were identified. EST-SSR distribution and characteristic analysis results showed that the mono-nucleotide repeats were the main repeat types, accounting for 41.0%. In addition, the sequences containing SSR were functionally annotated in Gene Ontology (GO) and KEGG pathway and were assigned to 49 GO categories, 242 KEGG pathways, among them 2 201 sequences were annotated against Nr database. By validating 235 EST-SSRs,74 primer pairs were ultimately proved to have high quality amplification. Subsequently, genetic diversity analysis, UPGMA cluster analysis, PCoA analysis and population structure analysis of 34 L. chuanxiong germplasm resources were carried out with 74 primer pairs. In both UPGMA tree and PCoA results, L. chuanxiong resources were clustered into two groups, which are believed to be partial related to their geographical distribution. In this study, EST-SSRs in L. chuanxiong was firstly identified, and newly developed molecular markers would contribute significantly to further genetic diversity study, the purity detection, gene mapping, and molecular breeding.

Objective To study the effects of environmental factors,including high temperature,high humidity and low temprtature,on the quality of whole blood.Methods Fresh blood was collected from 9 blood donors and divided into 40 parts.One group that comprised 20 samples was used to assess the effects of storage at high temperature and high humidity (0,3,6,12 and 24 h),and the other group(20 samples)was used to determine the effects of low temprtature(0,1,2 and 3 h).The serum free hemoglobin(FHb)concentration, plasma prothrombin time(PT), activated partial thromboplastin time(APTT),thrombin time(TT)and fibrinogen(FIB)were measured and analyzed.Results The high temperature(27-37℃)and humidity(60%-90%RH)had no effect on FHb, PT, APTT, TT or FIB of whole blood. The low temperature(0-13℃and -18--20℃)had some effect on the quality of whole blood.The FHb of test group was higher than that of control group.There was no significant difference in PT, APTT, TT and FIB between the two groups.Conclusion Under high temperature and humidity conditions, 24 hours of whole blood placement is feasible. Under low temperature or cold conditions,short-term whole blood placement is feasible in a tent with heating facilities.

OBJECTIVETo study the effects of hepatitis B virus (HBV) infection on the expression of Furin, an important proprotein convertase, in liver cells to provide insights towards its potential as a therapeutic target for improved antiviral efficacy.

METHODSFurin expression was measured in human liver specimens (infected tissues from patients with chronic HBV hepatitis vs. normal tissues from healthy donors) and in hepatoma cell lines (HBV-infected HepG2.2.15 cells vs. uninfected parental cell lines HepG2) using quantitative real-time RT-PCR (for mRNA), western blotting and immunohistochemistry (for protein).

RESULTSCompared to the uninfected tissues and cells, the HBV-infected tissue and cells showed down-regulated expression of furin at both the mRNA and protein levels. In particular, the HepG2.2.15 cells showed -50% less furin mRNA expression than the HepG2 cells and the difference was statistically significant (P less than 0.05).

CONCLUSIONHBV may suppress the host cell's expression of furin, possibly to benefit its survival and replication in the host cell.

Cell Line ; Furin ; metabolism ; Gene Expression Regulation ; Hep G2 Cells ; Hepatitis B virus ; physiology ; Hepatitis B, Chronic ; metabolism ; Host-Pathogen Interactions ; Humans ; Liver ; metabolism ; virology ; Proprotein Convertases ; metabolism ; Virus Replication

Objective To investigate the proliferative differences of adipose-derived stem cells (ADSCs) from neonatal suckling SD rats (5-d-old) and adult ones under the same culture condition.Methods ADSCs were isolated from the subcutaneous adipose tissues of neonatal suckling SD rats and adult ones,and then,type Ⅰ collagenase digestion was employed to obtain the ADSCs; the morphology of these cells was detected.The expressions of such cell surface markers as CD45,CD29 and CD90 were observed. The number of ADSCs on the 4th d of culture under the same condition and with the same planted density was compared between the neonate and adult rats. In vitro culture of the second generation of ADSCs was performed in the 96-well plates, and CCK-8 and alamar blue kit were employed to compare and quantitate the proliferative differences; optical density was observed by microplate reader. Results The ADSCs from neonatal SD rats and adult ones expressed the stem cell biomarkers: the expression of CD45 was negative, and that of CD29 was 98.04％ and 93.17％,respectively,and that of CD90 was 94.92％ and 93.3％,respectively,for neonate SD rat and adult ones.The cell counting results indicated that the number of ADSCs from neonatal rats ([8.87±0.13]×105 cells) was larger than that of adult ones ([4.51±0.36]×105 cells) after being cultured under the same condition and at the same planted density. The optical density value of ADSCs in neonatal rats was significantly higher than that in adult ones on the 6th and 7th d of culturing detected by CCK-8 kit and on the 2nd-7th d of culturing by alamar blue assay. Conclusion The proliferative ability of ADSCs from neonatal rats is greater than that of adult ones.

BackgroundGlaucoma is an optic neuropathy caused by structural damage of the optic nerve,and its early diagnosis is critical for arresting the irreversible damage of visual function. Optical coherence tomography (OCT) allows an early diagnosis of glaucoma by the measurements of the optic disc and retinal nerve fiber parameters. Objective This study was carried out to evaluate the effects of optic disc tomography and the measurement of the retinal nerve fiber layer (RNFL)thickness by spectral-domain OCT on the diagnosis of glaucomatous eye. MethodsIt was a noninterventional, cross-sectionalstudy. The optic disctopographic parameters and total and regional RNFL thickness were measured by RTVue OCT in 62 normal eyes and 67 glaucomatous eyes. The area under the receiver operating characteristic curve( ROC ) was used to assess the ability to differentiate glaucoma eyes from normal eyes of each testing parameter. This trial complied with the Helsinki Declaration and was approved by the Clinical Trial Ethic Committee of Beijing TongrenHospital. All of the participants signed the written informed consent before any medical examination. Results In the comparison of demography ,the ages of patients, the mean deficiency( MD ) and pattern standard difference( PSD ) of perimetry were obviously larger in the glaucoma group, primary open angle glaucoma ( POAG ) group and primary closure-angle glaucoma(PACG) group than those of normal controls( P＜0. 01 ). No significant differences were found in the disc area between a total glaucoma group, POAG group or PACG group and normal group ( P =0. 101,0. 741 and 0. 652, respectively) ;however, the average RNFL thickness between normal eyes and glaucomatous eyes were significantly different( 109. 758 μm versus 79. 539 μm, P＜0. 01 ). Among the eight regions around the optic disc, the thickest RNFL located at the inferotemporal( 150. 109 μm) and superotemporal( 146. 105 μm) regions in normal eyes,and at the superotemporal( 104. 354 μm) and inferotemporal( 102. 436 μm) regions in glaucomatous eyes. Both in normal and glaucomatous eyes,the thinnest RNFL located at the nasal(NU+NL) and temporal(TU + TL) regions. For optic disc topographic parameters,the highest ROC were observed in rim volume( ROC--0. 850,0. 841 and 0. 862 in total glaucoma,POAG and PACG, respectively) and vertical cup/disc ratio( ROC =0. 840,0. 849 and 0. 830 in total glaucoma,POAG and PACG,respectively), and the sensitivities for specificity cutoff set at 80％ were 73.1％ and 76. 1％ in total glaucoma,73.0％ and 81.1％ in POAG and 73.3％ and 70.0％ in PACG, respectively. For RNFL thickness ,the highest ROC was observed in average RNFL( ROC =0. 925,0. 910 and 0. 942 in total glaucoma, POAG and PACG,respectively) ,and the sensitivities for specificity cutoff set at 80％ were 89. 6％ ,89.2％ and 90. 0％ in total glaucoma,POAG and PACG, respectively. Among the eight regions around the optic disc, RNFL thickness of region IT achieved the highest ROC, RNFL thickness of region TU and TL had the lowest ROC. Conclusions RTVue OCT appears to be of fair discriminating ability in distinguishing normal from glaucomatous eyes. RTVue OCT shows promise for the diagnosis of glaucoma.

Objective To investigate the effect of nimesulide (NIM), a selective cyclooxygenase-2 (COX-2) inhibitor, on angiopoietins (Ang) gene expression of human glioma xenografts in nude mice and its significance. Methods Human SHG44 glioma cells were inoculated subcutaneously in 16 nude mice to establish xenograft models, and then these mouse models were randomly divided into NIM treatment group and control group. NIM (6 mg/kg) and saline were poured into the stomachs of the mice in each group, respectively, once daily for 35 d. The mRNA expressions of Ang-1 gene and Ang-2 gene in the xenografts were determined by RT-PCR. Microvessel density (MVD) in the xenografts was assessed by immunohistochemical technique. The tumor growth curve was drawn and the inhibition ratio of tumor growth was calculated. Results NIM could significantly inhibit the glioma xenografts growth with its inhibition rate reaching 42.03%. The mRNA expression of Ang-2 gene in NIM treatment group (0.2032±0.0185) was significantly lower than that in control group (0.6024±0.0289, P＜0.05), but that of Ang-1 gene showed no significant changes; therefore, the mRNA ratio of Ang-2/Ang-1 genes was decreased (0.5825±0.0621 vs. 1.5847±0.1948, P＜0.05). MVD in the xenografts of the NIM treatment group was significantly lower than that in the control group (P＜0.05). Conclusion NIM, by down-regulating the mRNA expression ofA ng-2 gene and changing the mRNA ratio of Ang-2/Ang-1 genes, can inhibit the tumor growth