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A new artificial somatic-autonomic neuroanastomosis has been established in male rats with spinal cord injury (SCI). Anorectal manometry and neural retrograde tracing were conducted in this animal model to analyze the mechanisms and the effects on recovery of anorectal function. The left L4 ventral root (L4VR) was intradurally micro-anastomosed to the L6 ventral root (L6VR) to establish the new regenerated neural pathway. Three months later the spinal cord was completely transected at the T9-10 level. Eight weeks later the model rats were randomly divided into two groups. The rats in the group 1 (n=8) were applied for anorectal manometry, and those in the group 2 (n=4) were used for neural retrograde tracing study with fluorogold (FG) and dextran tetramethylrhodamine (TMR). The results of anorectal manometry showed the new reflex pathway could induce rectum to contract and simultaneously electric activity of external anal sphincter (EAS) to become weak or disappearing (indicating synergetic relaxation of EAS). FG and TMR dual labeled neurons with round and elliptical shape were mainly observed in L4 angulus anterior of model rats. The regenerated neural pathways were effective to improve the rectum external sphincter synergetic status and restore the anorectal function.
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A protocol for the isolation, purification and culture of motor neurons from newborn rat spinal cord was described and the effect of glial cell line-derived neurotrophic factor (GDNF) on the growth of neurite of motor neurons was investigated in vitro. Spinal motor neurons (SMNs) were dissociated from ventral spinal cord of postnatal day 1 rats. The culture system for SMNs was established by density gradient centrifugation, differential adhesion, and use of serum-free defined media and addition of exogenous GDNF. After 72-h culture, the cells displayed the characteristic morphology of motor neurons, exhibited extensive neuritic processes and were positive for choline acetyltransferase (ChAT) expression. The neurite length of SMNs in GDNF groups was significantly longer than that in control group (P<0.05). This protocol can be adapted for various postnatal motor neurons studies.
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AIM:To investigate the activation and inactivation of nuclear factor kappa B(NF-κB)when tumor necrosis factor-related apoptosis-inducing ligand(TRAIL)is applied to induce the apoptosis of androgen-independent prostate cancer cell line PC-3M.METHODS:After the treatment of TRAIL or LPS at different doses,we tested the nuclear translocation of NF-κB by cell immunohistochemical staining and electrophoretie mobility shift assay(EMSA),and evaluated the level of IκB by RT-PCR under pyrrolidine dithiocarbamate(PDTC)treatment.RESULTS:EMSA and cell immunohistochemical analysis showed that the translocation of NF-κB was significantly activated when PC-3M cells were treated with TRAIL or LPS(P＜0.05).The pretreatment of PDTC upregulated the expression of IκB and blocked the nuclear translocation of NF-κB.CONCLUSION:TRAIL remarkably stimulates the activation of nuclear NF-κB in androgen-independent prostate cancer cells.On the other hand,the translocation of NF-κB can be significantly and efficiently inhibited in PC-3M cells by pretreatment with PDTC.The increased expression of IκB might be a clue for this inhibition,which means the possible way to enhance the effect of TRAIL in the apoptosis of prostate cancer cells.
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Connexin-43 (Cx43) expression in prostate cancer (PCa) cells and the potency of gap junctional intercellular communication (GJIC) in the cells were investigated, with an attempt to elucidate the reason why the so-called "bystander effect" mediated by thymidine kinase (TK) suicide gene therapy on PCa cells is not of significance and to explore the role of GJIC in PCa carcinogenesis.mRNA and protein expression of Cx43 in a PCa cell line PC-3m was detected by reverse-transcription polymerase chain reaction (RT-PCR) and strapt-avidin-biotin-enzyme complex (SABC) immunohistochemical staining, and inherent GJIC of PC-3m cells was assayed by scrape-loading and dye transfer (SLDT) assay. The expression of Cx43 in human normal and malignant prostate tissues was determined by SABC immunohistochemistry as well. It was found that Cx43 mRNA and protein expression in PC-3m cells was slightly reduced as compared with positive controls and the location of Cx43 protein was aberrant in cytoplasm rather than on membrane. Assessment of paraffin sections demonstrated that the expression of Cx43 protein in PCa cells was abnormally located and markedly diminished as compared with normal prostatic epithelial ones, displaying a negative correlation to the pathological grade (χ2=4.025, P＜0.05). Additionally, capacity of inherent GJIC in PC-3m cells was disrupted, which was semi-quantified as (+) or (-). It was indicated that both down-regulated expression of Cx43 mRNA and aberrant location of Cx43 protein participated in the mechanisms leading to deficient GJIC in PC-3m cells. Lack of efficient GJIC is a molecular event, which may contribute not only to limited extent of "bystander effect", but also to initiation and progression of prostatic neoplasm.
ABSTRACT
Connexin-43 (Cx43) expression in prostate cancer (PCa) cells and the potency of gap junctional intercellular communication (GJIC) in the cells were investigated, with an attempt to elucidate the reason why the so-called "bystander effect" mediated by thymidine kinase (TK) suicide gene therapy on PCa cells is not of significance and to explore the role of GJIC in PCa carcinogenesis. mRNA and protein expression of Cx43 in a PCa cell line PC-3m was detected by reverse-transcription polymerase chain reaction (RT-PCR) and strapt-avidin-biotin-enzyme complex (SABC) immunohistochemical staining, and inherent GJIC of PC-3m cells was assayed by scrape-loading and dye transfer (SLDT) assay. The expression of Cx43 in human normal and malignant prostate tissues was determined by SABC immunohistochemistry as well. It was found that Cx43 mRNA and protein expression in PC-3m cells was slightly reduced as compared with positive controls and the location of Cx43 protein was aberrant in cytoplasm rather than on membrane. Assessment of paraffin sections demonstrated that the expression of Cx43 protein in PCa cells was abnormally located and markedly diminished as compared with normal prostatic epithelial ones, displaying a negative correlation to the pathological grade (chi2=4.025, P<0.05). Additionally, capacity of inherent GJIC in PC-3m cells was disrupted, which was semi-quantified as (+) or (-). It was indicated that both down-regulated expression of Cx43 mRNA and aberrant location of Cx43 protein participated in the mechanisms leading to deficient GJIC in PC-3m cells. Lack of efficient GJIC is a molecular event, which may contribute not only to limited extent of "bystander effect", but also to initiation and progression of prostatic neoplasm.
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Summary: To study whether the sympathetic nerves coordinate with the parasympathetic nerves during micturition in the rat. We used antegrade neural tracing with biotinylated dextran amine (BDA) injected into the pontine micturition center (PMC) to label the terminals in the L6-S1 cord. Preganglionic parasympathetic neurons (PPNs) in the L6-S1 segment were labelled by retrograde transport of Fluorogold (FG) from the major pelvic ganglion (MPG).We detected retrograde neurons in L6-S1 using retrograde transport of horseradish peroxidase (HRP) from the intermediolateral cell column (IML) of the L1-L2 segment where sympathetic preganglionic neurons (SPNs) are located. Immunohistochemical methods showed that PPNs were identified to be choline acetyltransferase-immunoreactive (ChAT-IR). HRP-labelled neurons were not ChAT-IR and located dorsal to PPNs. BDA-labelled terminals were located mainly in the bilateral IML of L6-S1, some of which had synaptic contact with the HRP-labelled neurons. In addition, there were some wheat germ agglutinin-horseradish peroxidase (WGA-HRP) labelled terminals in the ipsilateral IML of the L1-L2 segment after WGA-HRP was microinjected into SPN. We conclude that PMC may control the preganglionic neurons of sympathetic nerves through the interneurons located dorsal to PPNs.
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The effectiveness and safety of ureteroscopic holmium: YAG laser lithotripsy for managing ureteral calculi was evaluated. Ureteroscopic holmium. YAG laser lithotripsy was performed in 168 ureteral calculi (upper 27 cases, middle 33 cases and lower 108 cases). The results showed that the stone-free rate was 92.6% in the upper ureteral calculi, 93.9% in the middle ureteral calculi and 94.4% in the lower ureteral calculi, respectively. The complication rate was 4.8% (8 cases). It was suggested that ureteroscopic holmium: YAG laser lithotripsy is a highly effective and safe treatment modality for managing ureteral calculi.
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Holmium , Lithotripsy, Laser , Methods , Ureteral Calculi , Therapeutics , UreteroscopyABSTRACT
The effectiveness and safety of ureteroscopic holmium: YAG laser lithotripsy for managing ureteral calculi was evaluated. Ureteroscopic holmium. YAG laser lithotripsy was performed in 168 ureteral calculi (upper 27 cases, middle 33 cases and lower 108 cases). The results showed that the stone-free rate was 92.6% in the upper ureteral calculi, 93.9% in the middle ureteral calculi and 94.4% in the lower ureteral calculi, respectively. The complication rate was 4.8% (8 cases). It was suggested that ureteroscopic holmium: YAG laser lithotripsy is a highly effective and safe treatment modality for managing ureteral calculi.
Subject(s)
Holmium , Lithotripsy, Laser/methods , Ureteral Calculi/therapy , UreteroscopyABSTRACT
To study the expression of beta-human chorionic gonadotropin (beta hCG) genes in renal cell carcinomas (RCC) and benign renal disease tissues, nested reverse transcription-polymerase chain reaction (RT-PCR) and restriction endonuclease analysis were employed to detect the expression of beta hCG genes in 44 cases of RCC tissues and 24 cases of benign renal disease tissues. It was found that 52% RCC samples revealed positive for beta hCG mRNA expression. Positive rate in advanced stage and poorly differentiated RCC was higher, but there was no significant difference. The positive rate of beta hCG mRNA expression was 54% in 24 cases of benign renal tissues, including 3 cases out of 6 polycystic kidneys, 7 cases out of 13 renal atrophies, 2 cases out of 2 oncocytomas and 1 case out of 2 pyonephrotic kidneys. beta 7 was most frequently transcribed subtype gene independent on the histology. These findings suggested beta hCG gene transcription is not only involved in RCC but also in benign renal diseases.
Subject(s)
Adult , Humans , Biomarkers, Tumor , Carcinoma, Renal Cell , Genetics , Metabolism , Chorionic Gonadotropin, beta Subunit, Human , Genetics , Kidney , Metabolism , Kidney Diseases , Genetics , Metabolism , Kidney Neoplasms , Genetics , Metabolism , Polycystic Kidney Diseases , Metabolism , RNA, Messenger , Genetics , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
Objective To study the changes of agrin expression in spinal cord motor neurons after somatic-autonomic ventral root(VR) anastomosis and somatic-somatic VR anastomosis,and the factors related to the synapse formation of foreign nerve regeneration. Methods Artifical somatic-automomic reflex pathway and somatic nerve self-anastomosis were established in rats.Retrograde tracing was carried out to label the motor neurons,and the neurons were isolated by flow cytometry.Real-time PCR was used to investigate the agrin mRNA expression of the neurons.Anterograde tracing and immunofluorescence technique were employed to label the new synapses of pelvic ganglia(PG). Results After 4 months of surgery,the somatic nerve can regenerate into PG successfully and form new synapses observed under laser scanning confocal microscope.The neurons involved in foreign nerve regeneration can be isolated successfully by retrograde tracing and flow cytometry.The expression of agrin in L_4-L_6 VRs anastomosis group was lower than that of the control group of L_4-L_4 VR self-anastomosis(P
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Objective To investigate the relationship between ?hCG genes expression and the clinicobiological behavior of bladder cancer. Methods RT-PCR method combined with restriction endonuclease analysis was adopted to detect the expressions of ?hCG mRNA and its subtypes in 37 cases of bladder cancer tissues. Results 26 of 37 cancer tissues (70%) were positive for ?hCG mRNA expression. The positive rate was higher in invasive tumor (T 2～T 4,13/14) compared with that in superficial tumor (Ta～T 1,13/23) ( P 0.05). Most of superficial tumors only expressed ?7 gene, while invasive tumor expressed ?7,?5, ?3, or ?8 . Conclusions The positive rate of ?hCG mRNA expression is related to tumor stage. The expression of hCG gene ?5, ?3 or ?8 in addition to ?7 indicates a worse differentiated or advanced bladder tumor.
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Objective To study the outcome of ureterocele trated by open surgery and by endoscopic manipulation. Methods We reviewed 29 cases of ureteroceles, including 16 intravesical ureteroceles and 13 extravesical ureteroceles. 19 cases were treated by open surgery and 10 by endoscopic procedure. Results 11 cases of intravesical ureteroceles and 8 cases of extravesical ureteroceles under went open surgery, The reoperation rate was 18.1% and 12.5%, respectively. 5 cases of intravesical ureteroceles and 5 cases of extravesical ureteroceles underwent endoscopic treatment,the reoperation rate being 40.0% and 80.0% respectively. Conclusions Endoscopic approach might be the primary management for intravesical ureteroceles, but open surgery is a favorable alternative for extravesical ureteroceles.
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Objective To study anorectal functional status in patients with lumbosacral spinal cord injury (SCI). Methods A total of 115 patients with lumbosacral SCI (SCI group) and 22 controls (control group) were studied by anorectal manometry. Results The maximal anal resting pressure was (56.7?31.1) mm Hg (1 mmHg=0.133 kPa) in SCI group and (87.2?29.2) mm Hg in control group, with significant statistical difference (P