DNA microarrays have been acknowledged to represent a promising approach for the detection of viral pathogens. However, the probes designed for current arrays could cover only part of the given viral variants, that could result in false-negative or ambiguous data. If all the variants are to be covered, the requirement for more probes would render much higher spot density and thus higher cost of the arrays. Here we have developed a new strategy for oligonucleotide probe design. Using type I human immunodeficiency virus (HIV-1) tat gene as an example, we designed the array probes and validated the optimized parameters in silico. Results show that the oligo number is significantly reduced comparing with the existing methods, while specificity and hybridization efficiency remain intact. The adoption of this method in reducing the oligo numbers could increase the detection capacity for DNA microarrays, and would significantly lower the manufacturing cost for making array chips.

OBJECTIVETo explore the molecular mechanism of APL cell resistance to ATRA.

METHODSThe ATRA sensitive and resistant APL cell lines, NB4 and NB4-R1, were used as in vitro models. The effects of specific inhibitors and activators of adenylate cyclase (AC) and phosphodiesterase (PDE) on ATRA-induced differentiation was evaluated by cell morphology, cell surface antigen expression and nitroblue-tetrazolium (NBT) reduction assays.

RESULTSSQ22536, a specific antagonist of AC, could dramatically block ATRA-induced NB4 cell differentiation. When ATRA + SQ22536 group compared with ATRA group, the positivity of CD11b decreased from (95.9 +/- 2.5)% to (60.3 +/- 7.1)%, while the A(540) in NBT reduction assay decreased from 0.585 +/- 0.092 to 0.170 +/- 0.028 (P < 0.05). Forskolin, an agonist of AC, could overcome the resistance of NB4-R1 cells to ATRA. When ATRA + forskolin group compared with ATRA group, the positivity of CD11b increased from (34.3 +/- 5.3)% to (94.6 +/- 2.4)%, while the A(540) in NBT reduction assay increased from 0.110 +/- 0.028 to 0.395 +/- 0.049 (P < 0.05). In contrast, the specific antagonist and agonist of PDE, 3-isobutyl-1-methylxanthine (IBMX) and calmodulin, exerted little impact on ATRA treatment.

CONCLUSIONSThe defaults in the initiation of AC activation may contribute to the resistance to ATRA in some APL cells.

Adenine ; analogs & derivatives ; pharmacology ; Adenylyl Cyclase Inhibitors ; Adenylyl Cyclases ; metabolism ; Antineoplastic Agents ; pharmacology ; CD11b Antigen ; metabolism ; Cell Differentiation ; drug effects ; Cell Line, Tumor ; Drug Resistance, Neoplasm ; drug effects ; Enzyme Activation ; drug effects ; Enzyme Inhibitors ; pharmacology ; Humans ; Leukemia, Promyelocytic, Acute ; metabolism ; pathology ; Phosphoric Diester Hydrolases ; metabolism ; Tretinoin ; pharmacology

OBJECTIVETo investigate the effect of Thymosin and growth hormone(GH) on inflammatory response in burn rats or burn rats with sepsis.

METHODSSixty-four SD rats were randomly divided into normal control group (NC, without treatment), sepsis group (S, with injection of LPS), sepsis + Thymosin group (ST, with successive injection of Thymosin and LPS), sepsis + GH group [SGH, with successive injection of recombinant human GH (rhGH) and LPS], burn group, burn + sepsis group (BS, with injection of LPS after burn), burn + sepsis + Thymosin group (BST, with successive injection of Thymosin and LPS after burn), burn + sepsis + GH (BSGH, with successive injection of rhGH and LPS after burn), with 8 rats in each group. Specimens of spleen tissues were harvested to determine HLA-DR in lymphocyte and evaluate inflammatory cell infiltration (score). Specimens of peripheral blood were collected to determine Toll-like receptor 4 (TLR4) level in monocyte and serum level of TNF-alpha, IL-4, IL-6, IL-10.

RESULTSCompared with those in NC group, serum level of IL-10 in S group decreased obviously, while other indices increased obviously (P < 0.01). The levels of HLA-DR and TLR4 and serum level of TNF-alpha were similar between SGH and ST groups (P > 0.05). Compared with those in SGH group [(2.87 +/- 0.04) score, and IL-6 (0.0083 +/- 0.0018) microg/mg, IL-4 (0.0102 +/- 0.0021) microg/mg, IL-10 (0.0310 +/- 0.0027) microg/mg, respectively], degree of inflammatory cell infiltration (1.50 +/- 0.76) score and serum levels of IL-6, IL-4, IL-10 of rats in ST group decreased obviously (0.0064 +/- 0.0012, 0.0058 +/- 0.0024, 0.0230 +/- 0.0021 microg/mg, respectively, P < 0.01). The levels of HLA-DR, TLR4 and inflammatory cell infiltration degree of spleen in B group were respectively higher than those in NC group and lower than those in BS group. Compared with those in NC group, serum levels of TNF-alpha, IL-6 in B group increased significantly, while IL-4, IL-10 showed an opposite tendency. There was no obvious difference between BST and BSGH groups in serum levels of HLA-DR and IL-6 (P > 0.05). Compared with those in BST group, inflammatory cell infiltration degree in spleen and the levels of TLR, TNF-alpha obviously decreased (P < 0.01), while IL-4 and IL-10 levels increased in BSGH group (P < 0.01).

CONCLUSIONSInhibitive effects between Thymosin and GH on extensive inflammatory reaction were similar with or without trauma, and GH has better effect as compared with Thymosin when with trauma.

Animals ; Anti-Inflammatory Agents ; pharmacology ; Burns ; immunology ; Human Growth Hormone ; pharmacology ; Inflammation ; immunology ; Male ; Rats ; Rats, Sprague-Dawley ; Sepsis ; immunology ; Thymosin ; pharmacology

OBJECTIVE: To study the clinical effect and safety of tacrolimus (TAC) combined with glucocorticoid (GC) versus mycophenolate mofetil (MMF) combined with GC in the treatment of primary IgA nephropathy (IgAN) in children. METHODS: A retrospective analysis was performed for the clinical data of children with primary IgAN confirmed by renal pathology between January 2012 and December 2017. These children were divided into TAC group and MMF group according to the treatment regimen. Their clinical data before treatment and at 1, 3, and 6 months of treatment were collected, and the remission status of IgAN and adverse reactions were compared between the two groups. RESULTS: A total of 43 children who met the inclusion criteria were enrolled, with 15 children in the TAC group and 28 children in the MMF group. At 1 month of treatment, there was no significant difference in the remission status between the two groups (P>0.05). At 3 and 6 months of treatment, the TAC group had a significantly better remission status than the MMF group (P<0.05). At 1 month of treatment, the TAC group had higher serum albumin levels than the MMF group (P<0.05). Both groups had a significant increase in serum albumin levels at each time point after treatment (P<0.0083) and a significant increase in the glomerular filtration rate (GFR) at 3 and 6 months of treatment (P<0.0083). There was no significant difference in the overall incidence rate of adverse reactions between the two groups (P>0.05), but fungal infection was observed in one child from the TAC group. CONCLUSIONS TAC combined with GC can effectively reduce urinary protein in children with primary IgAN, and it has a better short-term clinical effect than MMF combined with GC, with good safety.
Child ; Drug Therapy, Combination ; Glomerulonephritis, IGA ; drug therapy ; Glucocorticoids ; therapeutic use ; Humans ; Immunosuppressive Agents ; Mycophenolic Acid ; Retrospective Studies ; Tacrolimus ; therapeutic use

OBJECTIVETo study the growth and differentiation of adult canine autologous skeletal myoblasts after being transplanted into acute myocardial infarction (AMI) region by intramyocardium injection (IMI) and intracoronary infusion (ICI).

METHODSAutologous skeletal myoblasts were procured by a modified method. AMI model of adult canine was obtained through left anterior descending branch ligation and was divided into 4 groups (n = 5 for each group). Autologous skeletal myoblasts (1.0 - 1.4 x 10(8) cells) were injected locally into AMI region or infused into infarction-related coronary artery. Specimens were harvested 4 weeks after cellular transplantation for histological study including HE, PTH, immunochemical stain and transmission electronmicroscope.

RESULTSIn both two transplantation groups, newborn muscle-derived cells, striated muscle tissue and mature skeletal myofibril were demonstrated existing in MI region by electronmicroscope, PTH stain or anti-skeletal myosin heavy chain (slow) immunochemical stain, respectively. Newborn striated muscle tissues arranged in order of consistency with host myocardial fibers in two treatment groups. Newborn striated muscle tissue was more inclined to gather in MI region in the local injection group but distracted from each other in the intracoronary infusion group.

CONCLUSIONAutologous skeletal myoblasts appears to live and differentiate into mature striated muscle tissue after transplanting into AMI region by IMI or ICI routes.

Animals ; Cell Transplantation ; methods ; Cells, Cultured ; Dogs ; Female ; Male ; Myoblasts, Skeletal ; cytology ; transplantation ; Myocardial Infarction ; surgery ; Transplantation, Autologous

OBJECTIVETo assess the feasibility, safety and outcome of anatomical laparoscopic left lateral hepatic lobectomy for benign and malignant liver tumors.

METHODSFrom April 2005 to May 2008, 11 patients (7 male, 4 female; mean age 51.7 years) underwent anatomical laparoscopic left lateral hepatic lobectomy. Four patients presented with hepatocellular carcinoma and cirrhosis, while 1 patient had metastatic liver tumors from postoperatively colon cancer, five patients had hemangioma (2 cases with gallstones underwent cholecystectomy), 1 patient had a huge symptomatic angiolipoleiomyoma. Mean tumor size was 5.8 cm (range 2.1 to 12.0 cm). All the lesions were localized in the anatomical left lateral lobe (segments II to III).

RESULTSThe mean operative time was 147 min (range 120 to 180 min). There were no intraoperative or postoperative complications, and blood transfusions were not required. The mean postoperative hospital stay was 5.9 days.

CONCLUSIONSAnatomical laparoscopic left lateral hepatic lobectomy are feasible and safety.

Adult ; Aged ; Female ; Hepatectomy ; methods ; Humans ; Laparoscopy ; Liver Neoplasms ; pathology ; surgery ; Male ; Middle Aged ; Treatment Outcome

The present study focused on the potential mechanism of betulin (BT), a pentacyclic triterpenoid isolated from the bark of white birch (Betula pubescens), against chronic alcohol-induced lipid accumulation and metaflammation. AML-12 and RAW 264.7 cells were administered ethanol (EtOH), lipopolysaccharide (LPS) or BT. Male C57BL/6 mice were fed Lieber-DeCarli liquid diets containing 5% EtOH for 4 weeks, followed by single EtOH gavage on the last day and simultaneous treatment with BT (20 or 50 mg/ kg) by oral gavage once per day. In vitro, MTT showed that 0-25 mM EtOH and 0-25 μM BT had no toxic effect on AML-12 cells. BT could regulate sterolregulatory-element-binding protein 1 (SREBP1), lipin1/2, P2X7 receptor (P2X7r) and NOD-like receptor family, pyrin domains-containing protein 3 (NLRP3) expressions again EtOH-stimulation. Oil Red O staining also indicated that BT significantly reduced lipid accumulation in EtOH-stimulated AML-12 cells. Lipin1/2 deficiency indicated that BT might mediate lipin1/2 to regulate SREBP1 and P2X7r expression and further alleviate lipid accumulation and inflammation. In vivo, BT significantly alleviated histopathological changes, reduced serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) and triglyceride (TG) levels, and regulated lipin1/2, SREBP1, peroxisome proliferator activated receptor α/γ (PPARα/γ) and PGC-1α expression compared with the EtOH group. BT reduced the secretion of inflammatory factors and blocked the P2X7rNLRP3 signaling pathway. Collectively, BT attenuated lipid accumulation and metaflammation by regulating the lipin1/2-mediated P2X7r signaling pathway.

Aim To discuss the potential key mechanism of mogroside V in relieving pulmonary inflammation in asthmatic mice based on transcriptomics and proteomics. Methods Ovalbumin(OVA)was chosen to induce female BALB/C mouse asthma model, and the mice were treated with mogroside V to observe the pathological changes of lung tissues. Lung tissues in groups of natural control, ovalbumin control and mogroside V control were chosen for transcriptomic and proteomic analysis, and differential genes and proteins were screened for tendency analysis, followed by KEGG enrichment analysis for the potential genes and proteins. Results The results of lung morphological observation and HE revealed that mogroside V attenuated the OVA-induced pulmonary inflammation. Differential genes and proteins were selected from RNA-seq and DIA analysis. In the analysis of omics 454 genes increased in comparison between groups of natural control with ovalbumin control and decreased in comparison between groups of mogroside V control with ovalbumin control in 1 122 potential genes, and 111 genes were of opposite features. A total of 238 proteins increased in comparison between groups of natural control with ovalbumin control and decreased in comparison between groups of mogroside V control with ovalbumin control in 497 potential proteins, and 91 proteins were of opposite features. The PI3K/Akt signaling pathway was enriched from KEGG and tendency analysis of transcriptomics and proteomics. The key factors of Igha, Ighg1, PI3K and Akt increased in ovalbumin control group and decreased in mogroside V control group by the validation of molecualr biology experiments. Conclusions Transcriptomic and proteomic analysis exhibits that mogroside V relieves asthma in mice through inhibiting the activation of key factors including Lgha, Lgh1, PI3K and Akt, depressing the signaling pathway, attenuating pulmonary inflammation to reach the goal of moistening lung and relieve cough, which provides a reference for drug development of asthma.

Objective In emergency situations where simultaneous immunization by multiple vaccines are required,how to rapidly evaluate the effect of combined immunization is an urgent issue that needs to be solved.This study aimed to investigate the po-tential role and application value of the phenotypic changes of macrophages in rapid evaluation of the effect of combined Yersinia pestis and Brucella bovis vaccine immunization at early stage.Methods Y.pestis and B.bovis vaccines were injected into mice alone or in combination to establish animal models.The changes of the macrophage phenotypes(M1 or M2 polarization)and the CD8+T cell pheno-types and functions were detected in the early(4 d)and the late(14 d)stage of the immunization,respectively.The effect of the immuno-phenotype of macrophages at early stage on the function of CD8+T cells at late stage was analyzed.Results The co-immunization by Y.pestis and B.bovis vaccines led to the attenuation of the M1-polarization of macrophages at early stage,which were marked by de-creased expression of CD16/32 and increased expression of Detectin-1 on cell surface as well as decreased expression of IL-12 and in-creased expression of IL-4 inside the macrophage,in comparison with single vaccine groups,suggesting an interference between the two vaccines.Meanwhile,the activity of CD8+T cells(including the ratio of CD8+CD69+T,CD8+IFN-γ+T and CD8+GranzymeB+T cells) in combined immunization group showed similar tendency to the attenuated phenotypic M1-polarization of macrophages. Conclusion The phenotype of macrophages at the early stage of the co-immunization by Y.pestis and B.bovis vaccines showed consistency with the phenotype and function of CD8+T cells at late stage.It might give us some hint about the possibility of utilizing the phenotypic changes of macrophages to rapidly evaluate the effect of the co-immunization at early stage.

By preparing 10 batches of substance benchmarks freeze-drying powder( lyophilized powder),the methodology of the characteristic spectrum and the content of index component for substance benchmarks of Qingwei San was established. The characteristic peaks and the similarity range of the characteristic spectrum,the contents and the transfer rate range of isoferulic acid,palmatine and paeonol,and the paste-forming rate range were determined to define key quality attributes of substance benchmarks of Qingwei San. In the10 batches of substance benchmarks of Qingwei San,the similarity of characteristic spectrum was higher than 0. 90. In further comparison of the characteristic peak information,a total of 16 characteristic peaks were identified,including 5 characteristic peaks from Cimicifugae Rhizoma,5 characteristic peaks from Coptidis Rhizoma,2 characteristic peaks from Angelicae Sinensis Radix and 4 characteristic peaks from Moutan Cortex. The content of isoferulic acid was 0. 10%-0. 18%,with the average transfer rate of 49. 82%±4. 02%. The content of palmatine was 0. 17%-0. 31%,with the average transfer rate of 15. 84% ±2. 39%. The content of paeonol was 0. 41%-0. 75%,with the average transfer rate of 23. 41%±3. 23%. The paste-forming rate of the 10 batches of substance benchmarks were controlled at 27%-33%,with the transfer rate between the theoretical paste-forming rate and the actual paste-forming rate was 86. 59%±3. 39%. In this study,the quality value transfer of substance benchmarks of Qingwei San was analyzed by the combination of characteristic spectrum,the content of index component and the paste-forming rate. A scientific and stable evaluation method was preliminarily established,so as to provide the basis for subsequent development and quality control of relevant preparations of Qingwei San.
Benchmarking ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; Powders ; Quality Control ; Rhizome