Objective To observe the intluence ot chemokine RANTES influence on cardiac allograft acute rejection caused by alloreactive memory CD4+ T cells (Tm) adoptive transfer.Methods Heterotopic heart transplantation (HTx) from Balb/c donors to C57BL/6 recipients was performed by anastomosis of the vessels of the neck.Mice undergoing heterotopic heart transplantation received either adoptive transfer of 1 × 106 CD4+ Tm from the spleen of alloantigen-primed C57BL/6 mice or no cells (control group).After the cardiac transplantation,the mean survival time (MST),mean histologic rank of rejection,relative gene expression and serum concentration of RANTES in the cardiac grafts.Results (1) The percentage of CD4+ Tm was 26.83％ at the spleen of alloantigenprimed mice; (2) The MST was 5.17 ± 0.17 days in the CD4+ Tm+ HTx group versus 7.76 ± 0.21 days at the HTx group (control group) (P＜0.01); (3) The histological tests revealed that mean histologic rank of rejection activity in the sections of cardiac allografts on the day 5 post grafting was grade 3.92 ± 0.08 in the HTx+ CD4+Tm group versus grade 2.67 ± 0.14 in HTx group (P＜0.01) ;(4) The relative gene expression level of RANTES was 2.6 ± 0.21 in the CD4+ Tm + HTx group,significantly higher than in the control group (P＜0.01) ; (5) The serum concentration of RANTES in the CD4+ Tm+ HTx group was 223.6 ± 16.79 pg/mL,higher than in the control group (120.7 ±9.47 pg/mL,P＜0.01).Conclusion Alloreactive CD4+ Tm contribute to the increased expression and secretion of RANTES,and cardiac allograft acute rejection was more extensive in the CD4 + Tm + HTx group.

Objective To establish a zebrafish IGFBP-2 gene knock-down model by morphilino modified antisense oligonucleotide injection, so as to investigate the abnormal phenotypes of heart and vessels in early stage of zebrafish development and the expression of zebrafish cardiogenesis related genes. Methods The spatiotemporal expression of IGFBP-2 gene in early stage of zebrafish development was testified by whole mount in situ hybridization with antisense RNA IGFBP-2 probe. The IGFBP-2 morpholino (IGFBP-2 MO) that especially inhibited the gene promoter and standard control morpholino (Con-MO) were designed and synthesized by Gene-tools Corporation. Four different concentration gradients (0.05, 0.10, 0.25 and 1.0 mmol/L) were set as IGFBP-MO injection groups with 0.25 mmol/L Con-MO injection group and wild type group as controls. Contribution to the incidence of heart abnormal phenotypes and mortality rate induced by 4 different IGFBP-2 concentrations injection group was recorded and compared with 2 control groups. Heart abnormal phenotypes at different developmental stages in 0.25 mmol/L IGFBP-2 injection group were observed in detail. To validate the effectiveness of IGFBP-2 MO, the expression of enhanced green fluorescence presented by wild type zebrafish embryos at 12hpf which received single injection of IGFBP-2 EGFP recombinant plasmid and those co-injected with Con-MO or IGFBP-2 MO were detected. To investigate the regulation relationship between IGFBP-2 gene and other cardiogenesis related genes, expression of atrium specific marker gene Amhc was detected in IGFBP-2 MO and wild type group by in situ hybridization. Ventricle specific green fluorescence of Vmhc-EGFP transgenic zebrafish embryos whose IGFBP-2 gene was knocked-down were compared with those untreated. Zebrafish peripheral vascular development in the IGFBP-2 MO group was also checked out by micro-angiography. Results Whole mount in situ hybridization revealed that IGFBP-2 gene expressed in turn at eyes, midbrain and then focused on liver in early stage of zebrafish development. The micro-injection of 0.25 mmol/L IGFBP-2 MO resulted in heart malformation in nearly 60% of all injected zebrafish embryos. Heart malformation phenotypes included slow heart beat, pericardial edema, weak ventricle systole contraction and heart tube looping disorder. Some of them represented atria dilation, blood regurgitation and ciculation obstruction. Wild type zebrafish embryos that received single injection of IGFBP-2 EGFP plasmid DNA or co-injected with Con-MO presented strong enhanced green fluorescence at 12hpf, meanwhile, the fluorescence was barely seen in the embryos co-injected with IGFBP-2 MO. This strongly validated the gene specific knock-down effect of IGFBP-2 MO. Amhc was down-regulated at 48hpf in IGFBP-2 MO group. Vmhc-EGFP transgenic zebrafish down-regulated by IGFBP-2 gene also resulted in attenuated expression of ventriclar-specific green fluorescence protein at 48hpf. Intersegmental blood vessels of IGFBP-2 MO group by micro-angiography at 60hpf demonstrated an sparsate and chaos image, which suggested that IGFBP-2 gene expression was involved in the regulation of normal vascular development. Conclusions Micro-injection of IGFBP-2 MO is an efficient way to knock-down IGFBP-2 gene in zebrafish embryos. IGFBP-2 gene expression down-regulation leads to heart and vessels maldevelopment and have an impact on the expression of cardiogenesis related genes of zebrafish embryos as well. In short, IGFBP-2 plays a critical role in the normal cardiovascular development of zebrafish embryos.

OBJECTIVETo investigate the expressions of the EZH2 protein and EZH2 mRNA in human prostate cancer (PCa) and their correlation with the clinicopathologic parameters.

METHODSA tissue microarray (TMA) was constructed, which contained 48 dots of formalin-fixed paraffin-embedded tissue samples of human PCa. The expressions of the EZH2 protein and EZH2 mRNA in the samples were detected by immunohistochemistry (EnVision) and in situ hybridization (ISH). Another 15 cases of human benign prostate hyperplasia (BPH) and 12 cases of human prostate intraepithelial neoplasia (HGPIN) were taken as controls.

RESULTSThe positive rates of the EZH2 protein and mRNA were significantly higher in PCa than in BPH and HGPIN (87.5% vs 13.33% and 16.67%, 81.25% vs 6.67% and 16.67%, P < 0.05). The positive expression of the EZH2 protein was 96.67% and 72.22% in the Gleason score > or = 7 and Gleason score < or = 6 groups, respectively, with significant differences between the two groups (P < 0.05). The positivity of the EZH2 protein was significantly related to the TNM stage, increasing with tumor progression (P < 0.05), but not to age and serum PSA (P > 0.05), and so was that of EZH2 mRNA to TNM stage (P < 0.05), but not to age, serum PSA and Gleason score (P > 0.05). When the above characteristics were regarded as two-level discrete variables, both the EZH2 protein and EZH2 mRNA showed statistically significant differences in the positive expression rate (P < 0.05).

CONCLUSIONThe over-expressions of the EZH2 protein and EZH2 mRNA may play an important role in the pathogenesis and progression of PCa and provide some reference indexes for estimating the malignancy, progression and prognosis of PCa.

Aged ; Aged, 80 and over ; DNA-Binding Proteins ; genetics ; metabolism ; Enhancer of Zeste Homolog 2 Protein ; Humans ; Male ; Middle Aged ; Polycomb Repressive Complex 2 ; Prognosis ; Prostatic Hyperplasia ; metabolism ; pathology ; Prostatic Neoplasms ; metabolism ; pathology ; RNA, Messenger ; genetics ; Transcription Factors ; genetics ; metabolism

OBJECTIVEThis is a preliminary study of the endoscopic technique on open surgery. The purpose of this study is to introduce the use of endoscopic technique for the reconstruction of the condyle with costochondral graft through a preauricular incision.

METHODS33 patients of temporomandibular joint diseases (n=49) underwent reconstruction of mandibular condyle with costochondral graft. After preparation of the recipient site through a preauricular incision, maxillo-mandibular fixation, and preparation of the costochondral graft, the graft was fixed to the lateral side of the mandible ramus under the supervision of arthroscope.

RESULTSAll the patients had successful reconstruction of the condyle with costochondral graft without any severe bleeding or craniocerebral injury.

CONCLUSIONThis case series demonstrates the feasibility of endoscopic technique for the reconstruction of the condyle through a preauricular incision. It has the advantage of high efficiency, minimal postoperative morbidity, great patient comfort, and little appearance impairment. The endoscopic technique has a promising future.

Adult ; Arthroscopes ; Endoscopy ; Female ; Humans ; Male ; Mandible ; Mandibular Condyle ; Reconstructive Surgical Procedures ; Temporomandibular Joint Disorders

OBJECTIVETo investigate the expression of p120(ctn) in non-small cell lung cancer (NSCLC) and its correlation with the clinical and pathologic parameters.

METHODSImmunohistochemistry (S-P method) was used to detect the expression of p120(ctn) in 143 NSCLC cases. The variation of protein expression was further analyzed in 36 cases by Western blot. The correlation with clinical and pathologic parameters was studied.

RESULTSImmunohistochemically, normal bronchial cells showed membranous expression for p120(ctn), while NSCLC was characterized by cytoplasmic or diminished membranous staining. The rate of abnormal p120(ctn) expression was 79.7% (114/143). There was a significant correlation between abnormal expression of p120(ctn) and tumor differentiation, clinical stage, lymph node metastasis and poor prognosis (< 0.05), but not histologic typing. Western blot showed that the total amount of p120(ctn) in normal bronchial cells was significantly higher than that in NSCLC. The p120(ctn) isoform 1 (120,000) and isoform 3 (100,000) were expressed in normal lung tissue, while there was a reduced expression or absence of isoform 1 in NSCLC.

CONCLUSIONThe expression of p120(ctn) is abnormal in NSCLC; p120(ctn) may serve as a useful prognostic marker for NSCLC.

Aged ; Biomarkers, Tumor ; Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Catenins ; Cell Adhesion Molecules ; metabolism ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Lung ; metabolism ; pathology ; Lung Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; Phosphoproteins ; metabolism ; Prognosis ; Proportional Hazards Models

OBJECTIVETo investigate one-staged orthognathic surgery and arthroscope-assisted condylar reconstruction with costochondral graft for temporomandibular diseases combined with dental-maxillofacial deformities.

METHODSThe preoperative measurement and model surgery was performed. The patients underwent arthroscope assisted condylar reconstruction with costochondral graft combined with bimaxillary surgery or mandibular surgery.

RESULTSFrom Sept. 2003 to Dec. 2005, 15 cases were treated successfully with no bleeding, nerve injury and infection. The patients were followed up for 29 to 52 months (mean, 31.8 months). The appearance was markedly improved. TMJ function was good with no pain. The average maximum interincisal opening was 33.6 mm.

CONCLUSIONCompared with traditional operation, one-staged orthognathic surgery and arthroscope-assisted condylar reconstruction with costochondral graft has its advantages. Indication selection and preoperative design are very important.

Adult ; Cartilage ; transplantation ; Endoscopy ; Female ; Humans ; Male ; Mandibular Condyle ; surgery ; Reconstructive Surgical Procedures ; methods ; Ribs ; transplantation ; Temporomandibular Joint Disorders ; surgery ; Young Adult

Chronic ulcer of the lower extremities amounts for a grave and serious problem for public health. Western medicine focuses on controlling infection, improving blood circulation, surgical debridement, skin grafting, etc, but there are bottlenecks in the treatment. Traditional Chinese medicine (TCM) has a long history and a legacy of sound clinical efficacy in this area. TCM has developed a unique, effective external theory, and a large number of topical prescriptions and external technology. Through this research, a safe and effective treatment protocol of TCM for chronic ulcer of the lower extremities can be formed. To this end, during China's "Eleventh Five-Year" Plan, special research committees and projects on TCM external treatments and external technologies were established. This study on ulcer of the lower extremities constitutes one of the major research topics.

OBJECTIVETo investigate the necessity of modification to the traditional pigtail probe and evaluate its efficiency and therapeutic effect in searching the nasal cut ends and anastomosing the lacerated lacrimal canaliculus.

METHODSEighty-seven patients (including 87 eyes) suffering from canalicular laceration were randomized into two groups: 41 patients treated with traditional pigtail probes (Group A) and 46 with modified pigtail probes (Group B). During the reconstruction of the lacerated canaliculi, the traditional pigtail probe and the modified pigtail probe were used respectively to seek for the nasal cut ends of lacerated lacrimal canaliculi. Peripherally inserted central catheter (PICCTM) silicone tube with diameter of 0.95 mm was intubated as a stent for 4-6 months. The surgical outcomes were retrospectively analyzed after stent removal.

RESULTSIn Group B, the primary success rate of searching the nasal cut ends of lacerated lacrimal canaliculi was 93.48% (43/46) and the final success rate was 97.83% (45/46). No false passage formed in Group B. Statistical significance was found between Group A and Group B as the primary success rates of searching the nasal cut ends (X(2) equal to 10.522, P less than 0.01) and the false passage forming rates were concerned (X(2)) equal to 4.704, P less than 0.05), whereas no significance was found between the two groups as the final success rates were concerned (X(2) equal to 0.007, P larger than 0.05). The mean time of searching the nasal cut ends of lacerated lacrimal canaliculi in Group B was (5.02+/-0.73) minutes and the mean time of operation was (33.90+/-4.84) minutes, and both were significantly shorter than those of Group A (t(1) equal to 9.779, t(2) equal to 10.700, P less than 0.01). The cure rate of Group B was 95.65%, though higher than that of Group A, no statistical significance was found (Z equal to -0.007, P larger than 0.05). Totally, 2 patients (2.30%) were found to be absent of common canaliculus and underwent bicanalicular nasal intubation in the two groups.

CONCLUSIONSPigtail probes are efficient and convenient apparatus for searching the nasal cut ends of the lacerated lacrimal canaliculi in the reconstruction of canalicular laceration. Necessary or proper modifications to the pigtail probes can minimize the risk of iatrogenic damages or complications and enhance the efficiency and therapeutic effect of canalicular repair.

Adolescent ; Adult ; Anastomosis, Surgical ; instrumentation ; Equipment Design ; Eye Injuries ; surgery ; Female ; Humans ; Intubation ; instrumentation ; Lacerations ; surgery ; Lacrimal Apparatus ; injuries ; surgery ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; instrumentation ; Young Adult

OBJECTIVETo evaluate the hemodynamic response to passive leg raising (PLR) indicates fluid responsiveness in patients with septic shock.

METHODSTwenty patients with septic shock, considered for fluid challenge (FC), were enrolled in the study from June 2009 to May 2010. Hemodynamic changes were determined by pulse-contour derived cardiac index at baseline, before and after PLR, return to baseline for 10 min, before and after fluid challenge (250 ml saline for 10 min). An increase of SV after fluid challenge (FC-ΔSV) ≥ 10% were defined responders.

RESULTSTwenty patients with septic shock were included in the study. PLR and fluid challenge were performed 46 instances, among which 15 instances were defined as response group. SV and pulse pressure induced by PLR (PLR-ΔSV and PLR-ΔPP) were increased significantly in response group [(76 ± 19) ml vs. (65 ± 18) ml, (73 ± 20) mmHg vs. (62 ± 20) mmHg (1 mmHg = 0.133 kPa), P < 0.05], while in nonresponse group there were no significant change. PLR-ΔSV and PLR-ΔPP were correlated with FC-ΔSV (r = 0.51, P = 0.001; r = 0.45, P = 0.006), central venous pressure (CVP) were unrelated with FC-ΔSV. Area under curve (AUC) for PLR-ΔSV, PLR-ΔPP and stroke volume variation (SVV) were 0.846, 0.791 and 0.708. PLR-ΔSV ≥ 12.5% predicted fluid responsiveness with sensitivity of 80% and specificity of 93.5%. PLR-ΔPP ≥ 9.5% predicted fluid responsiveness with sensitivity of 73.3% and specificity of 83.9%.

CONCLUSIONSPLR-ΔSV and PLR-ΔPP can predict fluid responsiveness in patients with septic shock. PLR-ΔSV and PLR-ΔPP have a greater ability in predicting volume responsiveness than CVP and SVV.

Adult ; Aged ; Aged, 80 and over ; Female ; Hemodynamics ; physiology ; Humans ; Leg ; Male ; Middle Aged ; Posture ; Sensitivity and Specificity ; Shock, Septic ; physiopathology

OBJECTIVETo investigate the effect of rabbit saphenous and sciatic nerve homogenates on the proliferation and calcification of rabbit osteoblasts in vitro.

METHODThe saphenous nerves (sensory nerves) and the muscular branches of the sciatic nerve (motor nerve) were collected from 48 New Zealand white rabbits to prepare the nerve tissue homogenates. Bone marrow mesenchymal stem cells (MSCs) were isolated from the rabbits and cultured in vitro, and after 14 days of routine osteogenic induction, the resultant osteoblasts were identified by immunohistochemistry, alkaline phosphatase (ALP) and Alizarin red S staining. The osteoblasts were then incubated in the induction medium containing the saphenous (sensory nerve group) or sciatic homogenates (motor nerve group), with the cells in the dexamethasone-containing, dexamethasone-free osteogenic induction medium and control medium as the control. The proliferation, total protein and ALP activity of the osteoblasts were measured every other day until the 8th day, and Alizarin red S staining was used for quantitative analysis of calcification of the cells after two weeks.

RESULTSThe application of the saphenous nerve homogenates significantly promoted cell proliferation, total protein and ALP activity (P<0.01, P<0.05 and P<0.05), while exposure of the osteoblasts to dexamethasone inhibited the cell proliferation (P<0.001). Compared to dexamethasone-free group, the saphenous homogenates enhanced the mineralization of the osteoblasts (P<0.001).

CONCLUSIONSaphenous nerve homogenates significantly promotes the proliferation, differentiation, ALP activity and mineralization of rabbit osteoblasts, but sciatic nerve homogenates do not show osteogenic effects on the cells.

Animals ; Bone Marrow Cells ; cytology ; Calcification, Physiologic ; drug effects ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Culture Media ; pharmacology ; Female ; Male ; Mesenchymal Stromal Cells ; cytology ; Osteoblasts ; cytology ; physiology ; Rabbits ; Sciatic Nerve ; chemistry ; Sensory Receptor Cells ; chemistry ; Tissue Extracts ; pharmacology