Specific scAbs could be obtained through biopanning from phage antibody libraries by use of antigens as target molecules. scAbs specific to thrombin were separated from mouse antibody library by the panning strategy of alternating liquid-solid phase in this paper. Thrombin was biotinylated by photobiotin at first, then avidin-coated magnetic beads were utilized to isolate specific scAbs. The eluted phages were amplified and subject to the second round panning in microtiter plate to remove the unspecific reombinant phages. 4 specific scAbs were separated from 23 phage clones after four rounds of alternative panning.

Adolescent ; Adult ; Dermatologic Surgical Procedures ; Fascia ; injuries ; Fasciotomy ; Female ; Finger Injuries ; surgery ; Humans ; Male ; Skin ; injuries ; Soft Tissue Injuries ; surgery ; Surgical Flaps ; Young Adult

AIMThe structure of the human penile venous system has been well studied, but disappointing outcomes of penile venous surgery in certain patients have called into question on the anatomy. We planned to extend the anatomic knowledge with the ultimate goal of improving operative success.

METHODSThirty-five patients, who had undergone penile venous surgery, complained of poor erection developed gradually 6 months to 7 years postoperatively. Cavernosography was performed again during their return visit. Seven new patients underwent spongiosography followed by immediate cavernosography. Eleven male cadavers were carefully dissected. The anatomical findings were applied to venous surgery in 155 patients, who were then followed with the International Index of Erectile Function Questionnaire-5 (IIEF-5).

RESULTSImaging observation demonstrated that the deep dorsal vein served as a common vessel of the corpora cavernosa and corpus spongiosum. A prominent cavernosal vein was found coursing along each corpus cavernosum distally to the glans, in contrast to its reported description as a short segment at the penile hilum. All cadavers had two sets of para-arterial veins sandwiching the dorsal artery. In 148 men available for follow-up, their mean IIEF-5 score was 9.3 preoperative and increased to 22.7 after the operation. The 88.5% (131/148) of the patients believed that venous stripping was a worthy treatment modality. Five cases required sildenafil to maintain their potentia, which was not working preoperatively.

CONCLUSIONSThe failure of penile venous surgery has traditionally been ascribed to penile vein regeneration. However, our finding of a long and independent cavernosal vein and an independent set of para-arterial veins may be the principal cause in patients experiencing poor postoperative results.

Aged ; Dissection ; Erectile Dysfunction ; surgery ; Humans ; Male ; Penis ; blood supply ; diagnostic imaging ; surgery ; Phlebography ; Veins ; anatomy & histology ; surgery

Objective To investigate the influence of necroptosis pathways in microglia activation and inflammatory factor levels in cerebral ischemia/reperfusion (I/R) injury in rats and their significances.Methods A modified suture method was used to establish the models of middle cerebral artery I/R in rats.(1) SD rats according to the random number table were divided into cerebral I/R 6 h group,cerebral I/R 12 h group,cerebral I/R 24 h group,and cerebral I/R 72 h group (n=6);the expression of ionized calcium binding adaptor molecule-1 (iba-1) was detected by immunohistochemical staining,and time points enjoyed obvious iba-1 expression were selected according to the experimental results.(2) SD rats were randomly divided into sham operation group,cerebral I/R group,80 nmol necrostatin-1 (Nec-1) intervention group and 160 nmol Nec-1 intervention group (n=6),and the Nec-1 intervention was given 2 h after ischemia;Longa method was used to evaluate the neurological function scores and TTC method was used to detect the infarct volume;and the appropriate dosages of Nec-1 were selected according to these results.(3) SD rats were randomly divided into sham operation group,cerebral I/R group,solvent group and Nec-1 intervention group (n=6),and Nec-1 or DMSO intervention was given 2 h after ischemia.HE staining was used to observe the survival and proliferation ofmicroglias around the infarction tissues;immunohistochemical staining was used to detect the iba-1 expression surrounding the infarction tissues;immunohistochemistry and Western blotting were employed to observe the cytokine tumor necrosis factor (TNF)-α and interleukin (IL)-1β and glia-derived neurotrophic factor (GDNF) expressions.Results (1) The iba-1 expression at cerebral I/R 24 h group was significantly increased as compared with that in other groups (P＜0.05).(2) As compared with those in the cerebral I/R group,the neurological function scores and infarct volume were significantly decreased in the 80 nmol Nec-1 intervention group and 160 nmol Nec-1 intervention group (P＜0.05);more obvious changes were noted in the 160 nmol Nec-1 intervention group as compared with those in the 80 nmol Nec-1 intervention group,with significant difference (P＜0.05).(3) HE staining showed peri-infarct tissue inflammatory cell infiltration,reduced neuron number,cell body shrinkage and nuclear pyknosis in the cerebral I/R group,while these changes were less obvious in the Nec-1 intervention group;as compared with cerebral I/R group,the Nec-1 intervention group had significantly decreased expressions of iba-1,TNF-α,IL-1 β and GDNF (P＜0.05).Conclusion Necroptosis pathway involves in the activation ofmicroglias after I/R injury in rat brains;Nec-1 inhibits the activation of microglia,and reduces the neuronal damage by regulating inflammatory cytokines.

Female ; Hernia, Ventral ; surgery ; Humans ; Laparoscopy ; methods ; Middle Aged

OBJECTIVETo analyze the promoter methylation levels of p15, CDH1, DAPK and HICI genes of patients with myelodysplastic syndrome (MDS) and explore the relationship between the level of methylation and clinical features.

METHODSDNA methylation levels of p15, CDH1, DAPK and HICI in peripheral blood (PB) or bone marrow (BM) samples from 52 MDS patients were detected by real-time quantitative PCR. The correlation of the methylation level with clinical features and hematological findings was analyzed. 38 de novo AML patients and 46 normal individuals served as controls.

RESULTSThe methylation levels of p15, CDH1, DAPK and HICI were 16.23 ± 21.69, 6.59 ± 9.39, 0.14 ± 0.11 and 7.81 ± 9.70 in BM, and 14.96 ± 20.16, 6.00 ± 9.26, 0.12 ± 0.14 and 6.74 ± 9.72 in PB, respectively from 18 MDS patients, and the difference between BM and PB was not statistically significant (P > 0.05). The methylation levels of p15 (14.70 ± 18.17) and CDH1 (6.61 ± 8.79) genes in high risk (RAEBI/II) MDS were significantly higher than in low risk (RCMD/RARS/5q-, p15: 1.99 ± 1.59, CDH1: 1.23 ± 1.14 and RCMD, p15: 3.02 ± 3.42, CDH1:1.53 ± 2.06) MDS or control (p15: 1.69 ± 1.82, CDH1: 1.01 ± 1.12) (P < 0.05). The methylation levels of DAPK gene had no difference among subtypes of MDS, and that of HIC1 gene only differed between RAEB I/II (9.16 ± 11.95) and control (2.49 ± 2.26) (P = 0.042). The difference of methylation levels of p15, CDH1, DAPK and HICI in BM was statistically significant among subtypes of MDS (P = 0.001, 0.003, 0.039, 0.023, respectively). And so did of p15 and DAPK in PB (P = 0.013, 0.006, respectively). The methylation level of p15 and CDH1 was significantly correlated with IPSS classification and blasts percentage in BM.

CONCLUSIONSp15 and CDH1 genes are special hypermethylation genes in MDS. Methylation level of HIC1 gene showed an upward tendency from low risk to high risk MDS.

Adult ; Aged ; Aged, 80 and over ; Apoptosis Regulatory Proteins ; genetics ; metabolism ; Cadherins ; genetics ; metabolism ; Calcium-Calmodulin-Dependent Protein Kinases ; genetics ; metabolism ; Case-Control Studies ; Cyclin-Dependent Kinase Inhibitor p15 ; genetics ; metabolism ; DNA Methylation ; Death-Associated Protein Kinases ; Female ; Humans ; Kruppel-Like Transcription Factors ; genetics ; metabolism ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; metabolism ; Promoter Regions, Genetic ; Real-Time Polymerase Chain Reaction ; Young Adult

OBJECTIVE: To investigate the changes of phospho-JNK (p-JNK) during the incised wound healing of the skin in mice and to explore the rule of the time-dependent change of p-JNK in wound age determination. METHODS: The changes of p-JNK expression in incised skin wound were detected by immunohistochemistry and Western blot. RESULTS: There was a minimal baseline staining of p-JNK in control mouse skin. Changes of p-JNK expression were mainly detectable in neutrophils in the wound specimens from 3 hours to 12 hours after injury. Afterwards, the p-JNK positive cells were mostly mononuclear cells and fibroblasts between post-injury day 1 and day 5, whereas the p-JNK positive cells were mostly fibroblasts between post-injury day 7 and day 14. Morphometrically, the ratio of the p-JNK positive cells to the total increased gradually in the wound specimens from 3 hours to day 1, and maximized at day 1 with a slight decrease from post-injury day 3 to day 5. The ratio showed a second peak in the specimens of day 7, and then decreased gradually from post-injury day 10 to day 14. The changes of p-JNK expression were observed throughout the wound healing stages by Western blot as well, with a peak expression occurring between 12 hour and day 3 after injury. CONCLUSION p-JNK may play a pivotal role in inducing apoptosis of neutrophils, mononuclear cells, and fibroblasts during skin wound healing and meanwhile, p-JNK may be a potentially useful marker for wound age determination.
Animals ; Biomarkers ; Female ; Forensic Medicine ; JNK Mitogen-Activated Protein Kinases/metabolism* ; Male ; Mice ; Phosphorylation ; Random Allocation ; Skin/injuries* ; Time Factors ; Wound Healing ; Wounds, Penetrating/enzymology*

OBJECTIVE: To investigate the expression of cannabinoid receptor I (CB1R) during mice skin incised wound healing course and time-dependent changes of CB1R in wound age determination. METHODS: The changes of CBIR expression in skin incised wound were detected by immunohistochemistry and Western blotting. RESULTS: The control group showed a low expression of CB1R detected mainly in epidermis, hair follicles, sebaceous gland and dermomuscular layer. CB1R expression was undetectable in neutrophils in the wound specimens from 6h to 12h post-injury. CB1R positive cells were mostly mononuclear cells (MNCs) and fibroblastic cells (FBCs) from 1 d to 5 d post-injury. CB1R positive cells were mostly FBCs from 7 d to 14d post-injury. The ratio of the CB1R positive cells increased gradually in the wound specimens from 6 h to 3 d post-injury, reached peak level at 5 d, and then decreased gradually from 7d to 14 d post-injury. The positive bands of CB1R were observed in all time points of the wound healing course by Western blotting. The expression peak showed at 5 d post-injury. CONCLUSION CB1R is activated during the wound healing course. The expression of CB1R is found in mononuclear cells, which could be involved in inflammation reaction. CBIR is observed in fibroblastic cells, which could participate in the wound healing. CB1R may be a potentially useful marker for determination of wound healing age.
Animals ; Blotting, Western ; Disease Models, Animal ; Fibroblasts/metabolism* ; Forensic Pathology ; Immunohistochemistry ; Male ; Mice ; Monocytes/metabolism* ; Random Allocation ; Receptor, Cannabinoid, CB1/metabolism* ; Skin/metabolism* ; Staining and Labeling ; Time Factors ; Wound Healing ; Wounds and Injuries/pathology*

AIMTo report a series of varicocelectomy performed under pure local anesthesia.

METHODSFrom July 1988 to June 2003, a total of 575 patients, aged between 15 and 73 years, underwent high ligation of the internal spermatic vein for treatment of a varicocele testis under a regional block in which a precise injection of 0.8 % lidocaine solution was delivered to involved tissues after exact anatomical references were made. A 100-mm visual analog scale (VAS) was used to assess whether the pain level was acceptable.

RESULTSThe surgeries were bilateral in 52 cases, and unilateral in 523 cases. All were successfully performed on an outpatient basis except in the case of two patients, who were hospitalized because their surgeries required general anesthesia. Overall, 98.6 % (567/575) of men could go back to work by the end of the first post-operative week and only 8 (1.4 %) men reported feeling physical discomfort on the eighth day. The VAS scores varied from 11 mm to 41 mm with an average of (18.5+/-11.3) mm that was regarded as tolerable.

CONCLUSIONThis study has shown varicocelectomy under local anesthesia to be possible, simple, effective, reliable and reproducible, and a safe method with minimal complications. It offers the advantages of more privacy, lower morbidity, with no notable adverse effects resulting from anesthesia, and a more rapid return to regular physical activity with minor complications.

Acetaminophen ; administration & dosage ; Adolescent ; Adult ; Aged ; Analgesics, Non-Narcotic ; administration & dosage ; Anesthesia, Local ; Anesthetics, Local ; administration & dosage ; Follow-Up Studies ; Humans ; Lidocaine ; administration & dosage ; Male ; Middle Aged ; Outpatients ; Pain, Postoperative ; drug therapy ; Postoperative Complications ; Varicocele ; surgery ; Vascular Surgical Procedures ; methods

Objective:To investigate the uncoupling protein 2 (UCP2) expression and its relations with apoptosis, oxidative stress and mitochondrial division/fusion in HT22 cell model with intracerebral hemorrhage.Methods:HT22 cells cultured in vitro were divided into control group, and groups of 3, 6, 12 and 24 h after modeling; cells in the later 4 groups were given 10 μmol/L hemin for 3, 6, 12 and 24 h, respectively, and cells in the control group were given the same equivalent solvent for 24 h. The apoptosis of HT22 cells in these 5 groups was detected by Annexin V-FITC/propidium iodide Apoptosis Detection Kit. The reactive oxygen species (ROS) expressions in these 5 groups were detected by fluorescence probe method. Western blotting was used to detect the protein expressions of UCP2, apoptosis-related protein cleaved caspase 3, anti-apoptosis protein B cell lymphoma/leukemia2 (bcl2), and mitochondrial division/fusion proteins (Fis1, Drp1, Opa1, and Mfn2). Results:As compared with the control group, groups of 3, 6, 12 and 24 h after modeling had significantly increased apoptosis rate, ROS level, and UCP2, cleaved caspase 3, Fis1 and Drp1 expressions, and statistically decreased bcl2, Opa1, and Mfn2 expressions. Groups of 3, 6, 12 and 24 h after modeling had successively increased apoptosis rate, successively increased ROS levels, and successively increased UCP2, cleaved caspase 3, Fis1 and Drp1 expressions, and successively decreased bcl2, Opa1, and Mfn2 expressions, with significant differences ( P<0.05). Conclusion:UCP2 expresses in HT22 cell model with intracerebral hemorrhage in time-dependent manner, and it has close relations with apoptosis, oxidative stress, and dynamic balance of mitochondrial division/fusion of HT22 cells.