Objective To observe the expression of axon guidance cues Slit2 and Robo4 in lung tissue of rat with acute lung injury (ALI) and explore the function of Slit2 and Robo4 in ALI.Methods Forty-eight Sprague-Dawley rats were randomly (random number) divided into control group (n =24) and ALl group (n =24).ALI model was reproduced by cecum ligation and puncture (CLP).The control group only experienced a simulated operation without CLP.Both groups were further divided into 3 subgroups with 8 rats in each subgroup:12 h,24 h,and 48 h subgroups.artery blood gas analysis,lung tissue wet/dry weight (W/D) ratio,lung histopathologic changes,pulmonary microvascular permeability were observed.The serum tumor nocrosis factor-α (TNF-α) was measured with enzyme linked immunosorbent assay (ELISA).The expression of Slit2 and Robo4 mRNA were detected by reverse transcription-polymerase chain reaction (RT-PCR).The expression of Slit2 and Robo4 protein in lung tissues was assessed by immunohistochemistry.Date were analyzed by one-way ANOVA with SPSS version 13.0 software.Statistical significance was established at a P value of less than 0.05.Results Compared with the control group,in ALI rats at different time points,partial pressure of oxygen in arterial blood (PaO2) decreased significantly,lung W/D weight ratio and pulmonary microvascular permeability,the serum TNF-α increased significantly (all P ＜ 0.05),histopathology of lung revealed signs of injury.The expression of Slit2 mRNA in lung tissues was decreased markedly after CLP compared with control group [(0.56±0.13) vs.(0.87±0.05),F=41.39,P＜0.05,(0.42±0.10) vs.(0.85±0.07),F=93.54,P＜0.05,(0.26±0.08) vs.(0.89 ±0.09),F=227.05,P＜0.05].but there were no significant difference in expression of Robo4 mRNA in lung tissue between ALI group and control group [(0.86±0.07) vs.(0.83±0.05),F=0.695,P＞0.05,(0.82±0.05) vs.(0.89±0.08),F=2.061,P ＞ 0.05,(0.86 ± 0.08) vs.(0.86 ± 0.05),F =0.035,P ＞ 0.05].Immunohistochemistry study showed Slit2 protein was mainly expressed on the extracellular surface of vascular endothelial cells,while lung epithelial cell nuclei and endochylema.Robo4 protein was only expressed on the extracellular surface of vascular endothelial cells.Compared with the control group,expression of Slit2 protein in lung tissue in ALI group decreased markedly [(0.37 ± 0.05) vs.(0.45 ± 0.07),F =6.82,P ＜ 0.05,(0.32±0.06) vs.(0.47±0.09),F=23.54,P＜0.05,(0.28±0.07) vs.(0.46±0.06),F=28.01,P ＜ 0.05].As good as RT-PCR,there were no significant difference in expression of Robo4 protein in lung tissue between two groups [(0.53±0.04) vs.(0.52±0.05),F=0.155,P＞0.05,(0.53± 0.09) vs.(0.50±0.05),F=0.498,P＞0.05,(0.55±0.06) vs.(0.56±0.07),F=0.073,P ＞ 0.05].Conclusions Lung tissues of control group rats express Slit2 and Robo4.The decreased Slit2 mRNA and protein expressions in the lung tissue of rat with ALI caused by CLP may be associated with the occurrence of ALI.

OBJECTIVETo construct differential expression profiles of adenoid cystic carcinoma cell lines for screening candidate genes related to metastasis and to verify some candidate genes in adenoid cystic carcinoma.

METHODSRestriction fragments differential display PCR (RFDD-PCR) was used to set up gene expression profiles of adenoid cystic carcinoma cell lines-ACC-M and ACC-2, with high and low metastasis potential respectively. Candidate genes were screened through bioinformatics analysis. Then, a gene family of these candidate genes was checked using semi-quantitative reverse transcription-PCR(RT-PCR).

RESULTSTwo gene expression profiles including 5420 gene fragments were constructed, 12 genes of a family called matrix metalloproteinase genes (MMPs) were observed obvious differentially expressed between two cell lines. Results of semi-quantitative RT-PCR also identified this different expression of MMP2,MMP7,MMP9,MMP14,MMP15 and MMP24.

CONCLUSIONThe construction of gene expression profiles of ACC-M and ACC-2 cell lines makes the foundation for seeking the target genes of adenoid cystic carcinoma. MMP2,MMP7,MMP9 and MMP15 may be relevant with carcinogenesis, development and metastasis of adenoid cystic carcinoma, and different metastasis potential may result from different subtype of MMPs gene family.

Carcinoma, Adenoid Cystic ; enzymology ; genetics ; pathology ; Cell Line, Tumor ; Gene Expression Profiling ; Gene Expression Regulation, Enzymologic ; Gene Expression Regulation, Neoplastic ; Humans ; Matrix Metalloproteinase 15 ; genetics ; Matrix Metalloproteinase 2 ; genetics ; Matrix Metalloproteinase 7 ; genetics ; Matrix Metalloproteinase 9 ; genetics ; Matrix Metalloproteinases ; genetics ; Neoplasm Metastasis ; Reverse Transcriptase Polymerase Chain Reaction ; methods

Objective To observe the treatment results of 44 consecutive patients with refractory/relapse acute myeloid leukemia( AML) not in remission who received allogeneic hematopoietic stem cell transplantation ( allo-HSCT ) following decitabine (DAC)-intensified conditioning regimen (18) and idarubicin(IDA)-intensified conditioning regimen (26). Methods We conducted a retrospective study to evaluate the outcome of 44 consecutive patients with refractory/relapse AML not in remission who received allo-HSCT from 2009 July to 2016 May.Eighteen of them were given DAC-intensified conditioning regimen and 26 of them were given IDA-intensified conditioning regimen prior to allo-HSCT.The effects of DAC and IDA intensified conditioning regimen on the patients ' engraftment, transplant-related mortality, survival and occurrence of graft versus host disease(GVHD)were analyzed.Results and Conclusion In the DAC group, 7(38.9％) patients had acute GVHD (aGVHD).Among them,2 patients had grade Ⅰ aGVHD,5 patients had grade Ⅱ aGVHD.In the IDA group, 16(61.5％)patients had aGVHD.Among them,9 patients had grade Ⅰ aGVHD, 6 patients had grade Ⅱ aGVHD and 1 patient had grade ⅢaGVHD.In the IDA group, 9 of 26(34.6％) patients experienced leukemia relapse , all of them died due to the lack of effective therapies .In the DAC group, 4 of 18(22.2％) patients experienced leukemia relapse, 2 of them got long-term survival throughout salvage therapies .For the DAC group and IDA group , the 1-year probabilities of overall survival (OS) and leukemia-free survival (LFS) were 65.0％ versus 57.7％ (P=0.602) and 53.5％versus 57.7％(P=0.910), respectively.DAC-intensified conditioning regimen before allo-HSCT in the treatment of refractory/relapse AML is safe and effective.

Objective This study aims to investigate the structure and anti-angiogenic activity of homogeneous polysaccharides in Honeysuckle flowers,constructing the theoretical basis for its widespread application.Methods Homogeneous Honeysuckle polysaccharide LF-02-2 was obtained through water extraction,alcohol precipitation,anion exchange chromatography,and gel permeation chromatography.The structure of LF-02-2 was deduced through molecular weight determination,monosaccharide composition analysis,sugar residue linkage analysis,partial acid hydrolysis,and sugar aldonic acid reduction combined with NMR data.Additionally,in vitro tube formation experiments using human microvascular endothelial cells(HMEC-1)were conducted to assess its anti-angiogenic activity.Results Structural analysis of LF-02-2 revealed a homogeneous polysaccharide with a weight-average molecular weight of 74.1 kDa.The monosaccharide composition included rhamnose(Rha),galactose(Gal),galacturonic acid(GalA),and arabinose(Ara)in molar ratios of 10.43:14.94:6.66:67.97.The main chain consisted of 1,4-linked α-Galp A,1,2-linked α-Rhap,and 1,2,4-linked α-Rhap.Branches were connected to O-4 of 1,2,4-linked α-Rhap and included β-Galp with terminal connections and 1,4,6-linked β-Galp and α-Araf with terminal and 1,5-linked connections.Tube formation experiments demonstrated that LF-02-2 significantly inhibited tube formation in human microvascular endothelial cells(HMEC).Conclusion LF-02-2,an RG-I pectic polysaccharide from Honeysuckle flowers,exhibited significant anti-angiogenic activity,suggesting its potential for development as an anti-angiogenic drug.

Objective::To explore the effect of modified Qingyitang combined with continuous blood purification in the adjuvant treatment of severe acute pancreatitis (SAP) complicated with multiple organ dysfunction (MODS) caused by heat accumulation of viscera. Method::Totally 100 cases of patients of SAP complicated with MODS, who were diagnosed as heat accumulation of viscera by traditional Chinese medicine(TCM) and treated in ICU of the First Affiliated Hospital of Hunan University of Chinese Medicine during May 2015 and May 2019, were randomly divided into two groups, namely control group and observation group, with 50 cases in each group. The patients in control group were treated with fasting and abstinence, gastrointestinal decompression, inhibition of trypsin secretion, gastric mucosal protection, early jejunal nutrition, reduction of inflammatory reaction, continuous blood purification (CBP), mechanical ventilation and circulatory support. The patients in observation group were treated by nasojejunal tube according to syndrome differentiation in addition to routine comprehensive therapy. Modified Qingyitang was injected for 7 days. The remission time of abdominal pain and distention, the time of first exhaust and defecation, the time of ICU residence, the number of samples falling off, the cause of death and the number of cases were recorded. Relevant indexes were measured before treatment, on the 3^rd and 7^th day of treatment, including the evaluation indexes of pancreatitis: blood amylase (AMS), blood lipase (LPS), and modified computed tomography severity index (MCTSI), inflammatory response indexes were interleukin-6 (IL-6) and hypersensitive C-reactive protein (hs-CRP). Organ function indexes included APACHE-Ⅱ, arterial partial pressure of oxygen (PaO₂), oxygenation index (PaO₂/FiO₂), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), glutamyltransferase (γ-GGT), urine volume, creatinine (CREA), urea nitrogen (UREA), glomerular filtration rate (GFR), creatine kinase (CK), creatine kinase isoenzymes (CKMB), lactate dehydrogenase (LDH), myoglobin (Mb). Tissue perfusion evaluation indexes included acute physiology and chronic health score, serum lactic acid (Lac) and central venous pressure (CVP). TCM treatment score was based on the syndrome score of acute pancreatitis with heat accumulation of viscera syndrome. Result::The total effective rate of TCM syndromes was 86.67%(39/45) in observation group and 73.91%(34/46) in control group (χ² =13.524, P<0.01). On the 7^th day of treatment, the symptoms and indicators of the two groups were improved. Compared with before treatment, AMS, LPS, IL-6, hs-CRP, MCTSI, APACHE-Ⅱ, Lac, CVP, PaO₂, PaO₂/FiO₂, ALT and AST were improved on the 3^rd and 7^th day after treatment in observation group and control group. The levels of AMS, LPS, IL-6, hs-CRP, MCTSI, APACHE-Ⅱ, Lac, CVP, PaO₂, PaO₂/FiO₂, ALT, AST, ALP, γ-GGT, urine volume were significantly improved (P<0.05). Compared with control group on the 3^rd and 7^th day, the levels of AMS, LPS, IL-6, hs-CRP, MCTSI, APACHE-Ⅱ, Lac, CVP, PaO₂, PaO₂/FiO₂, ALT, AST, ALP, γ-GGT, urine volume were significantly improved (P<0.05). CREA, UREA, GFR, CK, CKMB, LDH and Mb were significantly improved (P<0.05). Compared with control group, the abdominal pain, abdominal distension relief time, first exhaust/defecation time, ICU stay time in observation group were significantly shortened (P<0.05), and the mortality rate in observation group was significantly reduced (P<0.05). Conclusion::Patients of SAP accompanied with MODS can be treated with blood purification combined with modified Qingyitang by promoting pancreas repair, inhibiting inflammation and improving organ function. It plays an important role in improving symptoms, alleviating TCM syndromes, delaying progression of disease, reducing hospital stay and reducing mortality.