Betulinic acid (BA) is a naturally occurring pentacyclic triterpene that is widely distributed throughout the plant kingdom. A variety of pharmacological activities have been ascribed to BA, including antitumor, anti-HIV, immunomodulatory, anti-oxidative, antimicrobial, anti-inflammatory, and antihelmintic activities. BA is a very promising new chemotherapeutic agent for the treatment of HIV infection and cancer due to the greater potencies and better therapeutic indices than some clinically used drugs. Notably, BA may play an increasingly important role in immune regulation and against oxidative damage as an immunomodulator and anti-oxidant. The pharmacological activities of BA are reviewed which provide a reference for the future research and utilization of BA.

Objective To investigate the expression of VIT-1 gene in melanocytes of patients with vitiligo, and to analyze the difference of its sequence. Methods The skin from the foreskins of healthy men by circumcision and from the non-lesional area on the buttocks of 5 patients were digested by dispase, then the epidermis and dermis were separated, and the melanocytes were isolated. Then we cultured the melanocytes from the controls in TICVA medium and those from the patients in TICVA medium supplemented with basic fibroblast growth factor (bFGF) and endothelin-1 ( ET-1). The expression of VIT-1 gene was measured by RT-PCR, the full-length cDNA of VIT-1 ORF was cloned and sequenced, and sequence difference was analyzed by CLUSTAL W ( 1.83 ) software. Results The expression levels of VIT-1 gene were significantly lower in melanocytes from the patients than in those from the controls. An 81 bp-intron was found in the VIT-1 ORF. VIT-1 was a fragment of FBXO11, located at its 3' end. Conclusion VIT-1 gene is not a new gene, but a fragment of FBXO11, and a member of F-box protein family.

The objective of this study was to explore the immunomodulatory effects of betulinic acid (BA) extracted from the bark of white birch on mice. Female mice were orally administered BA for 14 days in doses of 0, 0.25, 0.5, and 1 mg/kg body weight. We found that BA significantly enhanced the thymus and spleen indices, and stimulated lymphocyte proliferation induced by Concanavalin A and lipopolysaccharide as shown by MTT assay. Flow cytometry revealed that BA increased the percentage of CD4+ cells in thymus as well as the percentage of CD19+ and the ratios of CD4+/CD8+ in spleen. BA increased the number of plaque-forming cell and macrophage phagocytic activity as indicated by a neutral red dye uptake assay, and the peritoneal macrophages levels of TNF-alpha were also increased. In contrast, serum levels of IgG and IgM and serum concentrations of IL-2 and IL-6 were significantly decreased in BA-treated mice compared to the control as assayed by haemagglutination tests and ELISA, respectively. Taken together, these results suggest that BA enhances mouse cellular immunity, humoral immunity, and activity of macrophages. Thus, BA is a potential immune stimulator and may strengthen the immune response of its host.
Adaptive Immunity/*drug effects ; Animals ; Betula/*chemistry ; Cell Proliferation/drug effects ; Cytokines/blood ; Female ; Immunity, Innate/*drug effects ; Immunologic Factors/*pharmacology ; Macrophages/drug effects ; Mice ; Phagocytosis/drug effects ; Random Allocation ; Spleen/cytology ; Thymus Gland/cytology ; Triterpenes/*pharmacology

OBJECTIVERotavirus is the single most common cause of severe dehydrating diarrhea in young children worldwide, but the pathogenesis and immunity against this disease are not completely understood. A prospective study was conducted to assess gene expression of toll-like receptors (TLR) in children with acute rotavirus diarrhea.

METHODSSeventy-five children with acute rotavirus diarrhea and 38 control children were enrolled in this study from Sep. 2004 to Jan. 2005. All the 75 patients had detailed records of clinical characteristics. Rotavirus antigen was detected by ELISA from stools. Peripheral blood mononuclear cells (PBMC) were separated by Ficoll reagent and RNA was extracted by Trizol. The levels of mRNA for five TLRs in PBMC were examined by fluorescent quantitative RT-PCR.

RESULTSPatients with acute rotavirus infection had elevated mean levels of TLR 2, 3, 4, 7, 8 mRNA expressions in PBMC within 3 days since onset of the disease, P less than 0.05. But only TLR 2, 3, 8 mRNA levels remained increased in patients within 7 or 14 days since onset (P less than 0.05). Mean levels of mRNA for TLR 4 in PBMC was higher in patients with more severe diarrhea including longer duration of diarrhea, more episodes of diarrhea per day and higher severity scores (P less than 0.05).

CONCLUSIONManifold TLR may play roles in the start-up and regulation of immune responses in children with acute rotavirus diarrhea. These findings will be helpful to further recognize immune response in Chinese children with rotavirus diarrhea and, consequently, may provide directions and insights that could prove critical to the prevention or treatment of this important disease.

Acute Disease ; Antigens, Viral ; analysis ; Child, Preschool ; Diarrhea ; genetics ; virology ; Enzyme-Linked Immunosorbent Assay ; Feces ; virology ; Gene Expression ; Host-Pathogen Interactions ; Humans ; Infant ; Leukocytes, Mononuclear ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Rotavirus ; immunology ; physiology ; Rotavirus Infections ; genetics ; virology ; Toll-Like Receptors ; genetics

Accurate segmentation of multiple gliomas from multimodal MRI is a prerequisite for many precision medical procedures. To effectively use the characteristics of glioma MRI and im-prove the segmentation accuracy, we proposes a multi-Dice loss function structure and used pre-experiments to select the good hyperparameters (i.e. data dimension, image fusion step, and the implementation of loss function) to construct a 3D full convolution DenseNet-based image feature learning network. This study included 274 segmented training sets of glioma MRI and 110 test sets without segmentation. After grayscale normalization of the image, the 3D image block was extracted as a network input, and the network output used the image block fusion method to obtain the final segmentation result. The proposed structure improved the accuracy of glioma segmentation compared to a general structure. In the on-line assessment of the open BraTS2015 data set, the Dice values for the entire tumor area, tumor core area, and enhanced tumor area were 0.85, 0.71, and 0.63, respectively.

OBJECTIVE: To evaluate the effect of intrathecal pumping tramadol on cell-mediated immunity in rats with formalin inflammatory pain. METHODS: Thirty-two Sprague-Dawley adult male rats weighting 250 approximately 300 g were randomly divided into 4 groups (n=8 in each group):Saline group (NS) and 3 tramadol groups (T1,T2,and T3). The rats were anesthetized with intraperitoneal chloral hydrate (300 approximately 350)mg/kg. Microspinal catheter was inserted into the subarachnoid space at the lumber region according to modified Yaksh techniques. In the tramadol groups,after 5 days tramadol was continuously infused through the spinal catheter at 50 (T1),25 (T2), and 12.5 microg/h (T3) for 7 days. In the NS group normal saline was continuously infused instead of tramadol. On Day 7 formalin (5%, 50 microL) was injected into the plantar surface of the left hindpaw. The number of flinches, lickings and total time of licking was recorded for 60 min.Pain intensity scoring(PIS)(0 approximately 3;0= no pain, 3=severe pain) was used to assess the antinociceptive effect of intrathecal tramadol. The rats were killed after the evaluation of pain intensity. Body weight and spleen weight were measured and spleen index (spleen weight/body weight) was calculated. T-lymphocyte function was evaluated based on Concanavalin-A(ConA) induced splenocyte proliferation. A modified lactic acid dehydrogenase(LDH) release assay was done to assess the NK cell activity. Phenotypic expressions of cell surface markers of T lymphocyte subsets (CD3+, CD3+ CD4+, CD3+ CD8+, and CD4+/ CD8+) and NK cell(CD161+) in the spleen were analyzed by flow cytometry. RESULTS: The PIS scores were significantly lower in the T1,T2,and T3 groups than those in the NS group. The spleen index and splenocyte proliferation induced by ConA were significantly suppressed in the T1 group,and the phenotypes of T lymphocyte subsets were significantly changed,but no significant difference was found in the T2 and T3 groups compared with the NS group. There were no differences in NK cell activity in the 3 tramadol groups from the control group. CONCLUSION Intrathecal pumping tramadol has significantly antinociceptive effect. Intrathecal pumping higher dosage tramadol (50microg/h) suppresses T lymphocyte proliferation and alteres T lymphocyte subset phenotype but does not affect NK cell activity. General analgesic dosage tramadol (25 and 12.5 microg/h) has no effect on the immune function.
Analgesics, Opioid ; pharmacology ; Animals ; Dose-Response Relationship, Drug ; Formaldehyde ; Injections, Spinal ; Killer Cells, Natural ; immunology ; Male ; Pain ; chemically induced ; immunology ; Pain Measurement ; drug effects ; Rats ; Rats, Sprague-Dawley ; T-Lymphocyte Subsets ; immunology ; Tramadol ; administration & dosage ; pharmacology

OBJECTIVETo study the relationship between knee osteoarthritis (OA) and intercondylar notch narrowing based on the notch width index.

METHODSMagnetic resonance (MR) images were collected from middle-aged and elderly patients with a definite diagnosis of knee OA, including 42 with mild OA and 37 with moderate to severe OA, with 70 healthy individuals serving as the control group. The notch width indexes NWI, NWI-A, and NWI-P on the coronal images at different levels were calculated, and the intercondylar notch was classified, according to the features on axial MR images, into types A, U, and W. The association of OA with NWI, NWI-A, NWI-P, and notch type was determined, and the cutoff values were obtained based on the ROC curves at different levels as indicators for diagnosis of intercondylar notch stenosis.

RESULTSIn the control, mild OA, moderate to severe OA groups, the NWI value on coronal MR images were 0.252±0.019, 0.251±0.017, and 0.240±0.020, NWI-A were 0.261±0.024, 0.259±0.023, and 0.245±0.023, and NWI-P were 0.271±0.026, 0.270±0.024, and 0.254±0.022, respectively. Patients with moderate to severe OA had significantly smaller NWI, NWI-A, and NWI-P than the other two groups (P<0.05), and a significant association was found between NWI values at each level and the occurrence of moderate to severe OA (P<0.01). A NWI value<0.248, NWI-A<0.256, and NWI-P<0.266 supported a diagnosis of intercondylar notch narrowing. Type A intercondylar notch was found in the majority of patients with intercondylar notch narrowing (P<0.05).

CONCLUSIONPatients with moderate to severe OA have significant intercondylar notch narrowing, and patients with a type A intercondylar notch are more likely to have intercondylar notch narrowing than those with type U notch.

Aged ; Case-Control Studies ; Constriction, Pathologic ; Humans ; Knee Joint ; anatomy & histology ; Magnetic Resonance Imaging ; Middle Aged ; Osteoarthritis, Knee ; pathology ; ROC Curve

OBJECTIVETo analyze the relationship between the reference values of fibrinogen (FIB) in healthy Chinese adults and geographical factors to provide scientific evidences for establishing the uniform standard.

METHODSThe reference values of FIB of 10701 Chinese healthy adults from 103 cities were collected to investigate their relationship with 18 geographical factors including spatial index, terrain index, climate index, and soil index. Geographical factors that significantly correlated with the reference values were selected for constructing the BP neural network model. The spatial distribution map of the reference value of FIB of healthy Chinese adults was fitted by disjunctive kriging interpolation. We used the 5-layer neural network and selected 2000 times of training covering 11 hidden layers to build the simulation rule for simulating the relationship between FIB and geographical environmental factors using the MATLAB software.

RESULTSs The reference value of FIB in healthy Chinese adults was significantly correlated with the latitude, sunshine duration, annual average temperature, annual average relative humidity, annual precipitation, annual range of air temperature, average annual soil gravel content, and soil cation exchange capacity (silt). The artificial neural networks were created to analyze the simulation of the selected indicators of geographical factors. The spatial distribution map of the reference values of FIB in healthy Chinese adults showed a distribution pattern that FIB levels were higher in the South and lower in the North, and higher in the East and lower in the West.

CONCLUSIONWhen the geographical factors of a certain area are known, the reference values of FIB in healthy Chinese adults can be obtained by establishing the neural network mode or plotting the spatial distribution map.

Adult ; Asian Continental Ancestry Group ; China ; Climate ; Environment ; Fibrinogen ; Geography ; Humans ; Neural Networks (Computer) ; Reference Values ; Software ; Temperature

OBJECTIVETo investigate the antitumor effect of lycorine on renal cell carcinoma ACHN cells and explore the possible mechanism.

METHODSWe used flow cytometry to examine the effect of lycorine on ACHN cell cycle and apoptosis. The cell proliferation, migration and invasion were assessed with MTS assay, wound healing assay, and Transwell assay, respectively. Colony forming assay was performed, and the mRNA and protein levels of Bax, Bcl-2, survivin, caspase-3, cyclin D1 and CDK4 were measured with qRT-PCR and Western blotting.

RESULTSLycorine obviously inhibited the proliferation of ACHN cells with an IC(50) of 24.34 µmol/L. Lycorine also induced apoptosis of ACHN cells, caused cell cycle arrest at G(0)/G(1) phase, and suppressed the colony forming ability of the cells in a dose-dependent manner. The migration and invasion of ACHN cells were significantly inhibited by 5 µmol/L lycorine. Lycorine up-regulated the mRNA levels of CDK4, Bax, caspase-3 while down-regulated the levels of survivin, Bcl-2 and Cyclin D1; the protein levels of CDK4 and Bax were increased and cyclin D1, Bcl-2 and surviving expressions were decreased, but caspase-3 expression showed no significant changes following the treatment.

CONCLUSIONLycorine has obvious antitumor effect against ACHN cells, suggesting its value as a new therapeutic agent for renal cell carcinoma.

Amaryllidaceae Alkaloids ; pharmacology ; Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; Carcinoma, Renal Cell ; pathology ; Caspase 3 ; metabolism ; Cell Cycle Checkpoints ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; Cyclin D1 ; metabolism ; Cyclin-Dependent Kinase 4 ; metabolism ; Humans ; Inhibitor of Apoptosis Proteins ; metabolism ; Phenanthridines ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; bcl-2-Associated X Protein ; metabolism

Objective] To discuss the applicability of internal standard in determination of cycla-mate in wine by HPLC-tandem mass spectrometry . [ Methods] After dilution wine sample was deter-mined by HPLC-tandem mass spectrometry and quantified by external and internal standard methods respec-tively. [Results] The results by the 2 quantitation methods differed 11.2%in average, and the maxi-mum relative deviation was 16.2%.And the result by external standard quantitation method proved to be consistent with that by the international standard GB/T 5009.97-2003. [ Conclusion] The difference in results is thought to originate from matrix effect in HPLC-tandem mass spectrometry and the improper in-ternal standard compound applied .