Nucleotide-binding oligomerization domain protein 2 (NOD2) involves in host immune responses to pathogens and regulates natural immunity and specific immunity by identifying the peptidoglycan of bacterial cell walls and cleavage product muramyl dipeptide.As a newly discovered intracellular pattern recognition receptor,NOD2 plays important roles in the development of primary hepatocellular carcinoma by gene mutate,inducing liver inflammatory reaction and activating relevant signaling pathways.

Objective To investigate galectin3 on proliferation and migration of esophageal cancer Eca109 cells.Methods A lentiviral vector for over-expression of RNA targeting galectin3 was designed to transfect Eca109 cancer cells following plasmid-mediated transfection manual (Eca109/Gal3 cells).Inverted fluorescence microscope was used to observe the expression of EGFP.The proliferation of Eca109 cells was measured by cell counting Kit-8 assay.Eca109 cells apoptosis was determined by Annexin-V/7-AAD doublestaining.The migration capacity of Eca109 cells was determined in transwell assays.Western blot analysis was used to measure the expression of galectin3 protein.Results Galectin-3 expression was detected in Eca109 cells,with the Galectin3 expression in Eca109/Gal3 cells much more than non-transfected cells (t =14.33,P ＜ 0.05 ; t =10.28,P =0.037).Compared with non-transfected Eca109 cells,proliferation increased significantly in Eca109/Gal3 cells (t =-17.277,P ＜ 0.05 ; t =-13.4,P ＜ 0.05).Galectin3 evidently decreased in Eca109 cell apoptosis (t =3.053,P ＜ 0.05 ; t =5.446,P ＜ 0.05).Transwell migration assay showed that a greater number of Eca109/Gal3 cells crossed the artificial basement membrane compared with non-transfected Eca109 cells and negative control Eca109 cells (t =3.465,P ＜ 0.05; t =3.252,P ＜ 0.05).Conclusion Galectin3 expression is detected in transfected esophageal cancer Eca109 cells,whose overexpression can result in enhanced proliferation,migration,invasion as well as reduced apoptosis.These data indicate that in-depth research of galectin-3 may prove to be a potential molecular target for the treatment of esophageal cancer.

Objective To optimize the extraction process of volatile oil and inclusion preparation of hydroxypropyl-β-cyclodextrin from compoundJinling Sini Siwu Shixiaosan Granules.Methods The orthogonal approach of L9 (34) was applied to investigate the volume of volatile oil extraction by 3 factors to obtain the optimum extraction conditions. With inclusion rate of volatile oil and yield of inclusion compound as comprehensive evaluation indexes, the optimum preparation conditions were discussed. The characterization of this prepared inclusion was analyzed by TLC.Results The optimum extraction conditions were as follows:the crude medicine with medicinal powder was shattered into 20 meshes;8 times the amount of water was added and was extracted for 8 h. The best inclusion conditions were as follows:the proportion of volatile oil to HP-β-CD was 1∶6, with inclusion temperature of 30℃, grinding time of 2 h. Analysis results of TLC showed that stable inclusion compound was formed from volatile oil and HP-β-CD.Conclusion The optimized extraction process and inclusion preparation are with stable quality and high feasibility.

ObjectiveTo investigate the clinical characteristics in ABO-incompatible allogeneic peripheral blood stem cell transplantation(allo-PBSCT).Methods137 patients'clinical courses who accepted allo-PBSCT were retrospectively reviewed. Sixty-five cases of them were ABO-incompatible allo-PBSCT patients,including 32 ABO major mismatched cases,23 ABO minor mismatched cases and 10 ABO major and minor mismatched cases.Seventy-two ABO-identical cases were taken as control group.ResultsCompared with ABO-idential cases,the time of erythrocyte recovery after allo-PBSCT in ABO major and minor mismatched group was delayed [(73.2+10.3) d vs (97.5+10.4) d] (P ＜0.05).In ABO-incompatible group, the time of blood type switching in different ABO-incompatible types were found no significant difference (P ＞0.05) [ABO major mismatched:(45.7±17.3) d,ABO minor mismatched:(41.2+16.1) d and ABO major and minor mismatched:(48.4±20.9) d (P ＞ 0.05)].There were 8 cases who have a delayed time of blood type changing, including 6 cases demonstrated recipient-derived anti-A antibody. ConclusionABO-incompatible has no negative effect on allo-PBSCT.The time of erythrocyte reconstitution was delayed in ABO major and minor mismatched group.A delayed time of blood type switching tends to occur in ABO minor incompatible cases and patients who have anti-A antibody initially.

Objective To investigate the expression of nucleotide-binding oligomerization domain protein 2 (NOD2)in serum of patients with hepatocellular carcinoma (HCC),and to analyse the roles of NOD2 in HCC development and its clinical diagnostic value.Methods This study including 66 patients with HCC in the hospi-tal from March 1,2013 to December 31,2014 and 61 healthy controls.Serum NOD2 levels were determined by enzyme-linked immunosorbent assay (ELISA).Analysis of significance was performed with rank sum test using SPSS statistical 16.0 software.Results Serum levels of NOD2 in HCC patients were 171 pg/ml,significantly higher than that of healthy controls(95 pg/ml,Z =-5.00,P =0.00),and the serum NOD2 levels were correla-ted with clinical stage of HCC (H=56.26,P =0.00).Compared with the serum NOD2 levels in stageⅠ,Ⅱpatients (106 pg/ml)and healthy controls (95 pg/ml),the serum NOD2 level in stage Ⅲ and Ⅳ (220 pg/ml) were significantly increased (χ2 =31.24,P =0.00;χ2 =47.23,P =0.00),but the expression of NOD2 in stageⅠandⅡwere nearly equal to that of the healthy controls (χ2 =0.36,P =0.83).The ROC analysis revealed that the best diagnostic cutoff-point of serum NOD2 levels for predicting the Ⅲ and Ⅳ stages of HCC was 148.78 pg/ml,meanwhile corresponding sensitivity was 89.1% and specificity was 77.0%.Additionally,corre-lation analysis demonstrated that there was no significant correlation between NOD2 and alpha-fetal protein (r =0.44,P =0.14).Survival curves obtained that the survival time of HCC patients with NOD2 serum concentrations≥ 200 pg/ml was significantly less than that <200 pg/ml (χ2 =15.32,P <0.05).Conclusion NOD2 is highly expressed in the serum of HCC patients,especially in advanced patients,which is possibly involved in the development of HCC and has the potential to become an effective marker used for HCC diagnosis.

AIM: To investigate the expression of connective tissue growth factor (CTGF) and α-SMA in human lens epithelium cell (HLEC) line B3 after transfection by liposome-coated siRNA targeting CTGF.METHODS: HLECs were transfected with small interfering RNA (siRNA) targeting CTGF,labeled with 5`-fluorescein isothiocyanate (5`-FITC) and coated with lipofectamine.The transfection ratio was evaluated via fluorescence intensity.Cell counting kit-8 (CCK-8) assay was performed to assess cytoviability of both non-transfected and transfected HLECs.Quantitative RT-PCR,cell immunochemistry and Western blot analysis were conducted to detect the expression changes of CTGF and α-SMA after transfection.RESULTS: A highly effective transfection ratio was observed in siRNA co-transfected with lipofectamine.The transfection ratio reached 95% at 24h.The proliferation of HLECs was inhibited by siRNA after 72h transfection.The expression of CTGF and α-SMA significantly decreased in HLECs after transfected by CTGF siRNA for 24h.This effect was not found in negative control siRNA.CONCLUSIONS: SiRNA targeting CTGF decreased CTGF and α-SMA expression in HLECs,which is a potential therapeutic strategy for posterior capsular opacification.

OBJECTIVETo determine the outcome of combined anterior C2,3 reduction and fusion with posterior compressive C2 pedicle screw fixation for the management of unstable Hangman's fractures.

METHODSSixteen cases of unstable Hangman's fractures were retrospectively reviewed through X-ray, MRI and three dimensional CT scans. Pre- and postoperative radiographs were measured for translation and angulation of C2,3. Skull traction under extension poison was conducted in all the patients right after their admission. Then anterior C2,3 discectomy followed by interbody fusion, either with iliac autograft or with box cage, and locking plate fixation were performed in each case. Because dissatisfied reduction, mainly residual large fracture gap or kyphosis, was found by the C-arm fluoroscopy during operations, posterior compressive C2 pedicle screw fixation was performed in one stage. According to the Levine-Edwards classification, there were 12 cases of type II, 2 of type I a and 2 of type III in this group.

RESULTSFollow-up ranged 6-38 months, averaged 26 months. Fracture union and bone graft fusion were completed in an average of 4 months after operation. Complaints of neck pain and numbness of limbs disappeared in all patients after surgery, but range of neck motion decreased compared with normal people. Translation of C2 decreased from (4.2 +/- 1.4) mm preoperatively to (2.3 +/- 1.1) mm postoperatively, while angulation of C2,3 decreased from 8.6 degrees +/- 2.1 degrees preoperatively to 2.6 degrees +/- 1.0 degrees postoperatively. Both have statistical significance (P < 0.05). No implant failure or infection was observed.

CONCLUSIONSThe classification of Hangman's fracture should be modified in combination with MRI and CT scans to determine the stability of the fracture. Combined anterior C2,3 reduction and fusion with posterior compressive C2 pedicle screw fixation is the treatment of choice for patients with unstable Hangman's fractures.

Adult ; Aged ; Axis, Cervical Vertebra ; injuries ; surgery ; Bone Nails ; Cervical Vertebrae ; surgery ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; methods ; Humans ; Male ; Middle Aged ; Spinal Fractures ; surgery ; Spinal Fusion ; methods

Objective To explore the effect of patent foramen ovale(PFO)on white matter lesions(WMLs) in migraine without aura(MwoA).Methods Thirty-five patients with MwoA were examined by contrast transcranial Doppler(cTCD)and magnetic resonance imaging(MRI).According to the results of PFO and MRI Flair data,the patients' age,sex and headache characteristics were matched,and the WMLs were compared between the PFO positive group and negative group.Results Seven cases of WMLs were recruited in PFO positive group(19 cases)and the WMLs were distributed in the frontal lobe and/or the parietal lobe.The score ranged from 1 to 7 points.Five cases of WMLs were enrolled in PFO negative group(16 cases)and the WMLs also were distributed in the frontal lobe and/or the parietal lobe.The score ranged from 1 to 3 points.There was no significant difference in WMLs between the groups(P＞ 0.05).Conclusion White matter lesions in migraine without aura are distributed in the frontal lobe and the parietal lobe,and these findings do not support a relationship between PFO and WMLs.

BACKGROUNDProbucol is known to reduce the development of atherosclerotic lesions, but its impact on vascular remodeling associated with de novo atherosclerosis is incompletely understood. We therefore examined the effect of probucol on vascular remodeling in a rabbit model of established atherosclerosis.

METHODSAortic atherosclerosis was induced by a combination of endothelial injury and 10 weeks' atherogenic diet. Animals were then randomized to receive the foregoing diet without or with 1% (wt/wt) probucol for 16 weeks. At the end of week 26, in vivo intravascular ultrasound, pathological, immunohistochemical and gene expression studies were performed.

RESULTSProbucol significantly decreased vessel cross-sectional area, plaque area and plaque burden without effect on lumen area. More negative remodeling and less positive remodeling occurred in the abdominal aortas of probucol group than the control group (56% vs. 21%, 18% vs. 54%, respectively, both P < 0.01). In addition, the probucol group showed a smaller mean remodeling index relative to the control group (0.93 ± 0.13 vs. 1.05 ± 0.16, P < 0.01). Furthermore, probucol treatment decreased macrophage infiltration, inhibited apoptosis of cells within plaques, and reduced the production of matrix metalloproteinases-2, -9, cathepsin K and cathepsin S (all P < 0.01).

CONCLUSIONSThese findings suggest that probucol may attenuate the enlargement of atherosclerotic vessel walls and be associated with a negative remodeling pattern without affecting the lumen size. This effect may involve inhibition of extracellular matrix degradation and prevention of apoptosis in atherosclerotic plaques.

Animals ; Anticholesteremic Agents ; pharmacology ; Aorta ; pathology ; Apoptosis ; drug effects ; Atherosclerosis ; drug therapy ; metabolism ; pathology ; Lipids ; blood ; Macrophages ; drug effects ; physiology ; Male ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Oxidative Stress ; Probucol ; pharmacology ; Rabbits ; Ultrasonography, Interventional ; methods

OBJECTIVETo determine human epidermal growth factor receptor 2 (HER2) status in breast carcinoma by the techniques of a fully automated immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH), to compare the concordance of protein expression with gene amplification and to explore the optimization in process quality control.

METHODSA prospective study of invasive breast cancer specimens excised between May 2009 and April 2011 at the Cancer Hospital, Chinese Academy of Medical Sciences was conducted by automated IHC staining with the new 4B5 rabbit monoclonal antibody and FISH. An evaluation was performed according to the ASCO/CAP guidelines (2007) and Chinese guidelines (2009). The gene amplification status of 740 cases were detected by FISH.

RESULTSA total of 2420 cases of breast invasive ductal carcinoma without pre-operation therapy were tested by automated IHC. 551 cases (22.8%) were scored as positive (3+), 664 cases (27.4%) as equivocal (2+), and 1205 cases (49.8%) as negative (1+/0). Gene amplification was detected in 98.0% (242/247) HER2 protein expression positive (3+) cases and in 13.6% (53/389) equivocal (2+) cases. One of 247 (0.4%) HER2 expression 3+ cases and 5 of 389 (1.3%) HER2 expression 2+ cases were equivocal for gene amplification. No gene amplification was detected in expression negative (1+/0) cases by FISH (0/104). The overall concordance between IHC and FISH was 98.6% [(242 + 104)/(247 + 104)].

CONCLUSIONSThere is a high concordance rate between automated IHC with 4B5 rabbit monoclonal antibody and FISH results for assessing the HER2 gene amplification status in surgically-excised breast cancer specimens, suggesting that automated IHC with 4B5 antibody can provide a reliable method to detect HER2 overexpression for eligibility of HER2 targeted therapy.

Adult ; Aged ; Aged, 80 and over ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Carcinoma, Ductal, Breast ; genetics ; metabolism ; pathology ; Female ; Gene Amplification ; Gene Expression Regulation, Neoplastic ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Middle Aged ; Prospective Studies ; Quality Control ; Receptor, ErbB-2 ; genetics ; Young Adult