OBJECTIVETo investigate the association between transforming growth factor beta-1 (TGF-beta1) gene polymorphism and chronic allograft nephropathy (CAN).

METHODSFifty patients with failed renal allografts and clinically and histopathologically confirmed CAN were enrolled in this study along with another 50 renal transplant recipients with normal graft function. The DNA extracted from whole blood of the patients was amplified with PCR with sequence-specific primers for determining TGF-beta1 genotypes (position +869, codon 10 and position +915, codon 25). According to documented descriptions, the patients were classified into high and moderate-to-low cytokine production genotypes. The distribution frequencies of high production genotypes was then compared between CAN and non-CAN groups. To eliminate interference in the analysis of the association between TGF-beta1 polymorphism and CAN, other possible risk factors for CAN were screened, including the patients' gender, age, HLA match, delayed graft function, acute rejection, immunosuppressive regimen, cytomegalovirus infection, hypertension, and high cholesterol.

RESULTSCAN patients showed significantly greater proportion of high cytokine production genotype than the non-CAN group [70% (35/50) vs 38% (19/50), Chi(2)=10.306, P=0.001). Of the screened risk factors for CAN, only acute rejection showed some difference between the two groups, but analysis after subgrouping according to acute rejection did not suggest its influence on CAN, which supports the result that the rate of high production genotype was significantly higher in CAN group than in the non-CAN group.

CONCLUSIONMost CAN patients have high TGF-beta1 production genotype, which might be a risk factor for CAN after renal transplantation. TGF-beta1 genotyping can be of value in predicting the risk of CAN after renal transplantation.

Adult ; Female ; Genetic Predisposition to Disease ; Graft Rejection ; genetics ; Humans ; Kidney Diseases ; genetics ; Kidney Transplantation ; adverse effects ; Male ; Middle Aged ; Polymorphism, Genetic ; Risk Factors ; Sequence Analysis, DNA ; Transforming Growth Factor beta1 ; genetics ; Transplantation, Homologous

OBJECTIVETo assess the effect of local and intravenous transplantation of adipose tissue-derived stem cells (ADSCs) in promoting soft tissue wound healing in rats.

METHODSADSCs isolated from the adipose tissues of SD rats were cultured in vitro, and the third-passage cells were identified for their capacity of multipotent differentiation. Eighteen SD rats with 1.8 cm² dorsal full-thickness soft tissue defects (0.5 cm deep) were randomized into 3 groups to receive injection of 3.0×10⁶ DiI-labeled ADSCs via the tail vein, local injection of the cells at the wound site, or injection of saline (control). The wound healing was evaluated on days 3, 7, 11, and 14 postoperatively. On day 24 after the injury, tissue samples at the wound site were collected for fluorescent microscopy and HE staining.

RESULTSThe ADSCs obtained were capable of adipogenic, osteogenic, and neurogenic differentiation in vitro. ADSCs transplantation significantly promoted wound healing as compared to the control group. Obvious wound contracture was observed in the local injection group on day 3 and in the intravenous injection group on day 7. Fluorescence microscopy revealed DiI-positive cells in the healing wound, and HE staining showed a greater tissue thickness at the wound in the two ADSCs transplantation groups. Compared to the control group, the two ADSCs transplantation groups showed more gland-like structures and better neovascularization at the wound.

CONCLUSIONADSCs can significantly promote wound healing in rats, and local injection of ADSCs allows more rapid and obvious wound healing than tail veil injection of the stem cells.

Adipocytes ; cytology ; transplantation ; Adipose Tissue ; cytology ; Animals ; Cell Differentiation ; Cells, Cultured ; Male ; Rats ; Rats, Sprague-Dawley ; Stem Cell Transplantation ; methods ; Stem Cells ; cytology ; Wound Healing

OBJECTIVETo investigate the effect of adipose stromal vascular fraction cells (SVFs) on the survival rate of fat transplantation.

METHODS0. 5 ml autologous fat tissue was mixed with: 1) Di-labeled autologous SVFs ( Group A); 2) Di-labeled autologous adipose-derived stem cells (ASCs) (Group B); 3)Complete DMEM (Group C). And then the mixture was injected randomly under the back skin of 14 rabbits. The transplanted fat tissue in three groups was harvested at 6 months after implantation. Wet weight of fat grafts was measured for macroscopic aspects. After HE staining, blood vessel density, viable adipocytes and fibrous proliferation were counted respectively for histological evaluation. Trace of DiI-labeled ASCs in vivo was detected by fluorescent microscope.

RESULTSThe wet weight of fat grafts in group A (291.0 +/- 72.1) mg and group B (269.3 +/- 67.3) mg was significantly higher than that in group C (177.8 +/- 60.0) mg, but the difference between Group A and Group B was not significant. Histological analysis revealed that the fat grafts in group A and B was consisted predominantly of adipose tissue with less fat necrosis and fibrosis, compared with the fat grafts in group C. The grafts in both group A and B had significantly higher capillary density than those in the control group. Part of vascular endothelial cells were observed to origin from ectogenic DiI-labeled SVFs and ASCs.

CONCLUSIONSThe autologous isolated SVFs has a similar effect as autologous cultured ASCs to improve the survival rate of fat transplantation. And the former is more practical and safe, indicating a wide clinical application in the future.

Adipose Tissue ; cytology ; transplantation ; Animals ; Cells, Cultured ; Graft Survival ; Rabbits ; Stromal Cells ; cytology

OBJECTIVETo investigate the feasibility of using adipose tissue derived stem cells (ASCs) to promote neovascularization and survival rate of free fat transplantation.

METHODSASCs were isolated from aspirates from human liposuction and cultured in vitro. The cells were incubated in adipogenic, osteogenic, and chondrogenic medium for 2-4 weeks to induce adipogenesis, osteogenesis and chondrogenesis, respectively. ASCs were labelled by DiI. ASCs (A group), Insulin (B group), Medium (C group) were respectively mixed with free fat graft from aspirates. The mixtures were injected subcutaneously at the three random points on the back of eighteen 4- 6-week-old nude mice. Transplanted fat tissue was harvested after 6 months. The grafts were assessed by morphological observation, HE staining and immunohistochemistry.

RESULTSASCs can be easily harvested from liposuction aspirates and differentiate into adipogenic, osteogenic, chondrogenic lineages. The wet weight of transplanted fat tissue in ASCs group was (165.97 +/- 5.51) mg, significantly higher than that in the insulin group (93.42 +/- 5.12) mg and control group (67.64 +/- 5.09) mg (P = 0.000). The rate of fibrosis and steatonecrosis in ASCs group was( 152.2 +/- 9.8)/10HF, significantly lower than that in the Insulin group (743.9 +/- 20.4)/10HF and control group (892.2 +/- 16.5)/10HF (P = 0.000). DiI labelled ASCs were found between adipocytes and in the connective tissue in free transplanted fat tissue, and some of these cells were immunopositive for antihuman CD31 and FITC, suggesting differentiation into vascular endothelial cells.

CONCLUSIONSASCs can differentiate into vascular endothelial cells and contribute to angiogenesis in free transplanted fat tissue. ASCs can increase the survival rate and decrease the rate of fibrosis and steatonecrosis of free transplanted fat tissue. These findings suggest that ASCs-assisted transplantation may be an ideal cell therapy.

Adipocytes ; cytology ; Adipose Tissue ; cytology ; transplantation ; Adult ; Animals ; Cell Differentiation ; Cells, Cultured ; Feasibility Studies ; Female ; Graft Survival ; Humans ; Mice ; Mice, Nude ; Neovascularization, Physiologic ; Stem Cells ; cytology ; Tissue Scaffolds

Objective To explore the changes of functional connectivity of anterior cingulate cortex (ACC) in online game addicts during the resting state,and to analyze the function of ACC in the pathogenesis of online game addiction from a perspective of resting-state functional connectivity.Methods Seventeen online game addicts treated in our hospital from March 2011 to October 2011 were recruited as addiction group and 17 healthy controls at the same period were recruited as HC group.The baseline characteristics of all 34 subjects were investigated and compared between the addiction group and the HC group.All fMRI data were preprocessed after a resting-state fMRI scan,and then,the left and right anterior cingulate cortexes were selected as regions of interest (ROIs) to calculate the linear correlation between the ACC and the entire brain to compare the differences between the online game addicts and normal controls.Results Obvious differences between the addiction group and HC group were noted in hours and days of online game using and degree of thirst to play online games (P＜0.05);within the functional connectivity of ACC during the resting state,in contrast to the controls,the online game addicts showed increased connectivity with posterior cingulate,medium cingulate,midbrain,nucleus accumbens and supplementary motor area,but reduced connectivity with prefrontal cortex,temporal lobe and occipital lobe (P＜0.05).Conclusion Altered functional connectivity of the ACC reflects the dysfunction in ACC of online game addicts,which may be linked to the forming and maintaining of the online game addiction.

OBJECTIVETo investigate the quality and spatial distribution features of semen and to evaluate the reproductive health of the males in the Chongqing section of the Three-Gorge Reservoir area.

METHODSWe collected semen samples by masturbation after 2 -7 days of abstinence from the men in Nan'an, Shapingba, Zhongxian, Wanzhou, Yunyang and Wushan of Chongqing, which are geographically and demographically representative of the Three-Gorge Reservoir area. We analyzed the semen quality of all the samples and evaluated the reproductive health of the men.

RESULTSThe mean value of the five semen parameters of the male subjects from the six districts was within the normal range, including semen volume, sperm concentration, total sperm count, rapid progressive motile sperm, and total motile sperm. Those from Shapingba, Yunyang and Zhongxian exhibited abnormal sperm motility. According to the WHO criteria, normal value of all the semen parameters was found in less than 50% of the semen samples from the six districts, in 47% of those from Yunyang, and only 16% of those from Wanzhou. Spatial distribution maps of the semen parameters revealed significant spatial differences in seminal quality among the six districts, the highest in Yunyang, and the lowest in Wanzhou and Wushan that are located in the middle and lower reaches of the Three-Gorge Reservoir area.

CONCLUSIONThe mean value of semen parameters was low in a large proportion of men in the Chongqing section of the Three-Gorge Reservoir area, with spatial differences along the Changjiang river.

Adult ; China ; Humans ; Male ; Semen ; Semen Analysis ; Sperm Count ; Sperm Motility

OBJECTIVETo study the effects of benzo(a)pyrene (BaP) on the cell cycle distribution and activities of mitogen-activated protein kinase (MAPK) signal molecules (ERK1/2, JNK1/2 and p38) in human embryo lung cells (HELF), and to investigate the relationship between alterations of MAPK protein phosphorylation and the cell cycle distributions.

METHODSThe phosphorylation of MAPK were induced by exposing HELF cells to BaP at 0.1, 0.5, 2.5 and 12.5 micromol/L. The phosphorylation and protein expression levels of ERK1/2, JNK1/2 and p38 were determined through western-blotting assay. And the flow cytometry assay was used to measure the cell cycle effects in HELF cells after treatment with 2.5 micromol/L BaP for 24 h.

RESULTSThe phosphorylation levels of ERK1/2, JNK1/2 and p38 were significantly increased through BaP exposure. In addition, the phosphorylation of these three MAPKs has similar alteration pattern. We found that exposure of cells to 2.5 microM of BaP for 24 h resulted in a decrease of G(0) and G(1) population by 11.9% (F = 41.38, P < 0.01) and an increase of S population by 17.2% (F = 68.13, P < 0.01). Three chemical inhibitors of MAPK (AG126, SP600125 and SB203580) could significantly inhibit the cell cycle alteration because of BaP treatment.

CONCLUSIONERK1/2, JNK1/2 and p38 could positively regulate the BaP independently induced cell cycle alterations.

Benzo(a)pyrene ; toxicity ; Cell Cycle ; drug effects ; Cells, Cultured ; Fibroblasts ; drug effects ; metabolism ; Humans ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Lung ; cytology ; embryology ; MAP Kinase Kinase 4 ; metabolism ; MAP Kinase Signaling System ; drug effects ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Mitogen-Activated Protein Kinase 8 ; metabolism ; Mitogen-Activated Protein Kinase 9 ; metabolism ; Signal Transduction ; drug effects ; p38 Mitogen-Activated Protein Kinases ; metabolism

OBJECTIVETo study the role of E2F1/4 pathway in vitamin C reversing benzo (a) pyrene [B (a) P]-induced changes of cell cycle in human embryo lung fibroblasts (HELF) and the relationship between E2F1 and cyclin D1/CDK4.

METHODSThe stable transfectants, HELF transfected with antisense cyclin D1 and antisense CDK4, were established to detect the relationship of signaling pathway. Cells were cultured and pretreated with vitamin C before stimulation with B (a) P for 24 hours. The expression levels of cyclin D1, CDK4, E2F1 and E2F4 were determined by Western blot and the band intensity was analysed as the relative value to control by using the Gel-Pro 3.0 software. Flow Cytometric Analysis was employed to detect the distributions of cell cycle.

RESULTSB (a) P significantly elevated the expression levels of cyclin D1, CDK4, E2F1 and E2F4 in HELF cells. Vitamin C decreased the expression levels of above proteins in B (a) P-stimulated HELF cells. The expression levels of these proteins in B (a) P-treated above transfectants were lower than those in B (a) P-treated HELF cells. The expression levels of above proteins with vitamin C combined with antisense cyclin D1 were decreased as compared to those with antisense cyclin D1 alone. B (a) P increased the percentage of S phase as compared to the controls [(41.1 +/- 0.2)% vs (33.5 +/- 3.2)%, P < 0.05]. Both vitamin C [(33.2 +/- 0.6)% vs (41.1 +/- 0.2)%, P < 0.05] and antisense cyclin D1 [(31.2 +/- 1.3)% vs (41.1 +/- 0.2)%, P < 0.05] suppressed the changes of cell cycle induced by B (a) P. Vitamin C combined with antisense CDK4 markedly suppressed B (a) P-induced changes of cell cycle as compared to those with antisense CDK4 alone.

CONCLUSIONVitamin C might reserve the B (a) P-induced changes of cell cycle via intracellular signaling pathway of cyclin D1-CDK4/E2F-1/4.

Ascorbic Acid ; pharmacology ; Benzo(a)pyrene ; antagonists & inhibitors ; toxicity ; Cell Cycle ; drug effects ; Cyclin D1 ; metabolism ; E2F1 Transcription Factor ; metabolism ; E2F4 Transcription Factor ; metabolism ; Humans ; Lung ; cytology ; embryology ; Signal Transduction

OBJECTIVETo evaluate the integrity of the resected mesentery specimen after total mesorectal excision (TME) for low rectal cancer using methylene blue perfusion via the superior rectal artery.

METHODSTwenty patients with low rectal cancer were randomly divided into the methylene blue group (n=10) and the control group (n=10). All the patients received TME and macroscopic examination of the mesorectal surface was performed to evaluate the quality of the surgical specimen. The methylene blue was injected into the specimen postoperatively via superior rectal artery.

RESULTSThe mesorectal surface of all the specimens was intact on macroscopic examination. However, after methylene blue perfusion, 2 specimens were found to be incomplete. The number of lymph nodes in the methylene blue group were significantly larger (17.3+/-2.4 vs 12.4+/-5.4, P=0.016).

CONCLUSIONSIntegrity evaluation of TME specimen is necessary. Methylene blue perfusion is a convenient and effective method to identify subtle incompleteness of specimen and can improve the detection of lymph node.

Adult ; Aged ; Digestive System Surgical Procedures ; Female ; Humans ; Infusions, Intra-Arterial ; Male ; Mesenteric Artery, Inferior ; Mesentery ; pathology ; surgery ; Methylene Blue ; Middle Aged ; Postoperative Period ; Prognosis ; Rectal Neoplasms ; surgery ; Rectum ; blood supply

OBJECTIVETo evaluate the bowel control of the anus-preserving operation for elderly patients over 75 years with low rectal cancer.

METHODSThirty-nine elderly patients over 75 years with low rectal carcinoma (4-7 cm from anal verge) were treated during the study period. The patients were divided into different groups according to the surgical procedures and anastomotic locations. The bowel control and patients satisfaction were compared.

RESULTSThe time of recovering normal defecation frequency was (9.8+/- 2.9) months. There were no differences in bowel control and anorectal manometric findings between the lower anastomosis group and super-lower anastomosis group, the lower anastomosis group and anorectal anastomosis group. The patients in anorectal anastomosis group displayed significantly better bowel control and anorectal manometric findings than those in the super-lower anastomosis group (P< 0.05). The time of recovering normal defecation frequency in colonic J-pouch-anal anastomosis group was (7.7+/- 1.7) months, shorter than (10.6+/- 2.8) months in direct anastomosis group (P< 0.01). The complication rate of I degree incontinence was 36.1%, but there was no difference between the two groups. The anorectal manometric findings were better in J-pouch-anal anastomosis group than those in direct anastomosis group (P< 0.05).

CONCLUSIONColonic J-pouch-anal anastomosis for lower rectal carcinoma can significantly improve the bowel control in a short term without increasing the complication rate.

Aged ; Aged, 80 and over ; Anal Canal ; surgery ; Anastomosis, Surgical ; Defecation ; Fecal Incontinence ; etiology ; Female ; Humans ; Male ; Postoperative Period ; Rectal Neoplasms ; physiopathology ; surgery