Child ; Humans ; Intestinal Polyps ; genetics ; Molecular Biology

Objective To transfect PCDNA3.0-HIF1-? into the endothelial progenitor cells(EPCs),in order to provide the foundation for exploring myocardial vascular repairing capacity and angiogenesis of the EPCs transfected with HIF gene.Methods The EPCs were separated by the method of density gradient separation in the human bone marrow,and then the cells were inoculated in the fibronectin-packaged petri dishes complete medium with the concentration of 1?106?mL-1,and cultivated for 14 d. The expressions of the endothelial cell-specific components ecNOS and flk-1 were detected with RT-PCR; the cells were stained with acLDL-Dil and FITC-UEA-1; the PCDNA3.0-HIF1-? plasmid was transfected into the EPCs mediated by LipofectamineTM 2000,the transcription of HIF1-? was determined with RT-PCR; the expression of VEGF was detected with enzyme-linked immunosorbent assay (ELISA); the expression of HIF1-? was detected with Western blotting.Results After cultivated for 5 d,some round monocytes transformated into spindled inherent EPCs;7 d later,there were some cell colonies containing dozens of cells;10 d later,there were obvious cell clones; RT-PCR showed ecNOS band in 548 bp,flk-1 band in 819 bp; acLDL-Dil and FITC-UEA-1 double-staining was positive; RT-PCR showed specific band in 383 bp; ELISA showed the content of VEGF in the supernatant of EPCs transfected with HIF1-? was (20.53?2.33) ?g?L-1,and it was (3.96?1.67) ?g?L-1 in the EPCs transfected without HIF1-?,there was significant difference between two groups (P

OBJECTIVETo investigate the role of endogenous hydrogen sulfide (H2S) in erectile dysfunction (ED) induced by androgen deficiency.

METHODSWe randomly divided 30 eight-week-old healthy male SD rats into six groups: 2-week control (A), 4-week control (B), 2-week castration (C), 4-week castration (D), 2-week castration + androgen replacement (E), and 4-week castration + androgen replacement (F), those in groups E and F subcutaneously injected with testosterone propionate (TP) at the physiological dose of 3 mg/kg per day after castration, while those in the other groups with isodose oil instead. At 2 and 4 weeks after operation, we determined the level of serum testosterone (T) , intracavernous pressure (ICP) , mean carotid arterial pressure (MAP) of the rats, measured the concentration of H2S in the plasma and corpus cavernosum tissue, and detected the expressions of cystathionine-P3-synthase (CBS) and cystathionine-gamma-lyase (CSE) by immunohistochemistry and Western blot.

RESULTSThe serum T level was significantly lower in group C ([0.63 +/- 0.15] nmol/L) than in A ( [ 16.55 +/- 4.17] nmol/L) and E ( [ 18.99 +/- 4.62] nmol/L) (P <0.05), as well as in group D ([0.70 +/-0.22] nmol/L) than in B ([15.44 +/-5.18] nmol/L) and F ([20.99 +/-6.41] nmol/L) (P <0. 05) , and so were ICP/MAP after 5 and 7 V electrical stimulation of the pelvic ganglia (P <0. 05) , H2 S concentration (P <0.05), and the expressions of CBS and CSE (P <0.05). The expressions of CBS and CSE proteins were also significantly decreased in group C as compared with D (P <0.05).

CONCLUSIONThe reduced expressions of CBS and CSE may inhibit the H2 S signaling pathway, which might be one of the mechanisms underlying androgen deficiency-induced ED in rats.

Androgens ; deficiency ; Animals ; Cystathionine beta-Synthase ; metabolism ; Cystathionine gamma-Lyase ; metabolism ; Erectile Dysfunction ; metabolism ; Hydrogen Sulfide ; metabolism ; Male ; Orchiectomy ; Penis ; metabolism ; Rats ; Rats, Sprague-Dawley

Objective To explore the influence of the lentiviral vectors-mediated mouse genetic engineering regulatory T cells (Treg) infused after allogeneic bone marrow transplantation (alloBMT) on graft-versus-host disease (GVHD) and graft-versus-leukemia (GVL) effect in mice.Methods Lentivirus-mediated expression of Forkhead box P3 (Foxp3) transformed CD4 + CD25- T cells from Balb/c mice into engineered Tregs in vitro. An allo-BMT model of Balb/c→C57BL/6 mice was established. The recipients were given lethal X-ray total body irradiation before transplantation.Mice were randomly assigned into five groups and each group contained 10 recipients: (1) The recipients in radiation group were injected with 0.2 ml RPMI 1640; (2) The recipients in leukemia control group were injected with 5 × 106 donor bone marrow cells and 500 mouse T-cell leukemia/lymphoma cells (EL4 cells); (3) The recipients in transplantation control group were injected with 5 × 106 donor bone marrow cells and 5 × 106 splenocytes plus 500 EL4 cells; (4) The recipients in engineering Treg group were injected with 5 × 106 donor bone marrow cells, 5 × 106 splenocytes and 500 EL4 cells plus 5 × 106 genetic engineering Treg; (5) The recipients in empty vector control group were injected with 5 × 106 donor bone marrow cells, 5 × 106 splenocytes and 500 EL4 cells plus 5 × 106 empty vector-transduced CD4+ CD25- T cells. Survival time, clinical GVHD score or histopathological analysis (skin, liver and small intestine) were observed after allo-BMT. Chimerism of bone marrow cells from recipients survived for 60 days after transplantation was measured. Results The mean survival time in radiation group, leukemia control group, transplantation control group,engineering Treg group and empty vector control group was ( 10. 3 ± 1.5), (20. 7 ± 1.9), (26. 0 ±4.3), (49. 0 ± 17. 7) and (24. 4 ± 4. 1 ) days respectively. The survival time in engineering Treg group was significantly prolonged as compared with other groups as judged by the log-rank test (P＜0. 05).Histopathological analysis in several target organs (skin, liver and small intestine) confirmed the presence of severe GVHD in transplantation control group and empty vector control group. No histological signs of GVHD or leukemia were observed in recipients in engineering Treg group and clinical GVHD scores in this group were significantly decreased as compared with transplantation control group and empty vector control group. Conclusion Co-injection of genetic engineering Treg can efficiently prevent recipients from lethal GVHD without affecting GVL activity during allo-BMT in mice.

AIM: To investigate whether or not short-term high pressur e proliferates human umbilical vein endothelial cells (HUVECs), and the role of pp60c-src in cell proliferation. METHODS: Cultured HUVECs of 3-6th passa ge were exposed to atmosphere 0 mmHg (AP), 120 mmHg (MP), 180 mmHg (HP) and interfered with cap top ril(Cap, 10 ?mol/L)or irbesartan(Irb, 10 ?mol/L) Cell proliferation was q uantified by measuring hexosaminidase activity. The content of pp 60 c-src in cells was investigated by Western blotting. RESULTS:① Hexos aminidase activities which reflected cell number increased significantly at 4 h in MP and HP(0 145?0 018 and 0 144?0 019 vs 0 118?0 003,P

OBJECTIVETo investigate the feasibility and the mid-term effects of unilateral pedicle screw fixation and transforaminal lumbar interbody fusion in treating lumbar degenerative diseases.

METHODSFrom August 2005 to May 2010, 56 patients with lumbar degenerative diseases underwent lumbar posterolateral fusion,their clinical data were retrospective analyzed. The patients were divided into two groups (unilateral group and bilateral group) according to fixation methods,27 patients in unilateral group who were underwent unilateral pedicle screw fixation, including 18 males and 9 females with a mean age of (57.5 ± 7.1) years old (ranged from 41 to 66 years); and 29 patients in bilateral group who were treated with bilateral pedicle screw fixation (on the basis of the above, with contralateral vertebral pedicle screw fixation), including 19 males and 10 females with a mean age of (54.6 ± 5.1) years old (ranged from 43 to 68 years). The clinical data such as operation time, blood loss volume, hospitalization time and cost were compared between two groups. JOA score system was used to evaluate the neurological function. And fusion status and cage-related complication were also analyzed.

RESULTSAll patients were followed up from 36 to 60 months with an average of 45.8 months. No iatrogenic nerve, blood vessels or organs injury were found during operation. Operation time, blood loss volume, hospitalization time and cost in unilateral group were better than that of bilateral group (P < 0.05). There was no significant difference in JOA score between two groups (P > 0.05). Two patients in unilateral group developed with cage related complications, 1 case was cage displacement and 1 case was cage subsidence, while 2 patients in bilateral group developed with complications of no-fusion, and there was no significant differences between two groups (P = 0.58).

CONCLUSIONUnilateral pedicle screw fixation is a satisfactory method and can obtain good effects in treating lumbar degenerative diseases in mid-term, however, the indications should be well considered.

Adult ; Aged ; Biomechanical Phenomena ; Female ; Humans ; Intervertebral Disc Degeneration ; physiopathology ; surgery ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Pedicle Screws ; Spinal Fusion ; methods

ObjectiveTo investigate the application of intraoperative Computed Tomograph (CT) using in surgery for complex acetabular fractures.MethodsFrom June 2008 to December 2010,14 patients (9 males,5 females; with the mean age of 45.1 years; range,28-62 years) with complex acetabular fractures were operated using intraoperative CT.Preoperative radiotherapy and CT scan were adopted to evaluate the fractures.Three dimensional reconstruction based on CT scan was used to mimic surgery.The surgery approach and the type of internal fixators were noted.Intraoperative C-arm and CT scan were used to evaluate the fractures reduction respectively.Decision of additional reduction was made by surgeons according to above mentioned methods respectively and the results were noted.Comparing to preoperative design,the change of surgery plan were noted.Overall time,frequency and radiation dose of intraoperative CT scan were also noted.ResultsAll patients in this study received average 2.7 times of intraoperative CT scan.Mean time of CT scan was 40.4 min and the overall dose of radiation was 47.2 mGy.Decision of additional reduction was made in 3 cases according to C-arm radiography and 4 cases according to CT scan (above mentioned 3 cases were included).The change of surgery plan was made in one case.In postoperative radiography evaluation according to Matta's score system,anatomical reduction were achieved in 8 cases,imperfect reduction in 3 cases and poor reduction in 3 cases.ConclusionIntraoperative CT scan increases the radiation time and dose of patients dramatically.When used to evaluate fracture reduction intraoperatively,it can't take the advantage of traditional C-arm radiography.When delicate preoperative plan is made with radiography and three dimensional reconstruction based on CT data,the efficiency of intraoperative CT scan for complex acetabular fractures are to be discussed.

Objective:To summarize our experience on perioperative anesthesia management of patients with crush syndrome caused by earthquake under field condition.Methods:The clinical anesthesia data of 10 patients with crush syndrome caused by China Wenchuan earthquake,who were treated under field condition,were retrospectively analyzed,and our experience on perioperative anesthesia management was summarized.Results:After volume therapy,urine alkalization,and stabilization of internal environment,the 10 patients underwent dehridement,decompression by fasciotomy or amputation under epidural anesthesia or brachial plexus block plus psychological therapy and moderate tranquillizer.During the operation,the patients showed smooth respiratory and cardiovascular characteristics without pain.Conclusion:Under field condition,regional anesthesia can provide adequate analgesic effect during and after the surgery.Early fluid resuscitation and life support are vital to the safety and life of patients.

Objective To investigate the association of angiotensinogen(AGT)gene A-6G、T174M and G-217A polymorphisms with the risk of essential hypertension(EH)in the elderly of Han nationality.Methods Genotypes of AGT gene A-6G,T174M and G-217A polymorphisms in 177 aged EH patients and 86 sex and age-matched controls were analyzed with gene chip technology.Results The A-6G and T174M polymorphisms of AGT gene were significantly associated with EH.The numbers of the three genotypes of A-6G were 113,58 and 6 in the patient group and 70,15 and 1 in the control group(P= 0.014)and those of T174M were 94,77 and 6,60,25 and 1(P=0.031),respectively.G-217A polymorphism was not related to EH.Individuals carrying A-6G AA and T174M CC genotypes showed 57% and 56% lower risk of EH(OR=0.43;95%CI=0.23-0.82 and OR=0.44;95%CI=0.25-0.79, respectively).Conclusions The A-6G AA and the T174M CC genotype may be related with decreased risk of EH and G-217A polymorphism may have little role in the etiology of EH in Han nationality.

Objective To improve the processing protocol of the frozen erythrocyte ( RBC) washing machine of the dialysis type to enhance its efficiency of treating the 2 U frozen RBCs .Methods The frozen RBC washing machine for dialysis was used to deglycerolize the 2 U frozen RBCs after thawing .Measures including adjustment of the Pump speed , washing buffer adding and solution osmolarity were taken to decrease the processing time .Hemoglobin(Hb) content, Hb recovery, residual glycerol, free Hb content, deforming ability, and apoptosis of the deglycerolized RBCs were assessed to evaluate the efficiency of the improved protocol .Results As for the cells processed respectively by the previous and improved protocols, contents of Hb were (35.03 ±4.20) g and (53.82 ±2.08) g, respectively; Hb recoveries were (61.10 ±8.46)% and (80.22 ±3.73)%,respectively.Osmolarities representing residual glycerol were (307.00 ± 14.50)mOsm and (322.00 ±29.00)mOsm.Free Hb values were (0.74 ±0.04) and (0.80 ±0.08)g/L.The processing time was (1.64 ±0.31) and (1.04 ±0.16) h,respectively.There were significant differences in Hb content , Hb recovery, and deforming ability between the cells processed by the two protocols respectively .However , there were no significant changes in glycerol contents , free Hb, and apoptosis extent between the cells processed by the two protocols .Conclusion The innovated protocol can improve the quality of washed frozen RBCs , decrease the processing time and enhance the efficiency of the machine.