Objective To study the relationship between the DNA content of chondrocytes in the costal cartilage and postmortem interval in putrefactive rat cadavers.Methods Nuclear DNA wag visualized by modified Feulgen's staining method.DNA content of ehondrocytes in the costal cartilage was semi-quantita tively determined by a computerized image analysis system in rats within 35d postmortem.Results Staining intensity of the nuclei was gradually reduced within from 1d to 28d postmortem.The nuclej could not be detected at 35d.The DNA content of chondrocytes decreased time-dependently within 28 days after death as determined semi-quantitatively,which revealed a linear relationship between DNA content and postmortem interval.Conclusion DNA content of chondrocytes in the costal cartilage reduces time-dependently with the extension of postmortem interval.

Objective To investigate the correlation between ~(18)F-fluorodeoxyglucose (FDG) uptake and hypoxia inducible factor1α (HIF-1α) level,microvessel density (MVD) in human gliomas.Methods ~(18)F-FDG PET scan was performed preoperatively in 41 patients with gliomas (including 23 highgrade and 18 low-grade tumors).The ratios of maximum standardized uptake value(SUV_(max))between tumor (T)and contralateral white matter (WM) were calculated (T/WM).Immunohistochemical stain methods were used to evaluate the level of HIF-1α and measure the MVD in tumors.Correlation analysis between SUV_(max) of T/WM and HIF-1α level,MVD wag performed.The t-test,one-way ANOVA test,Spearman rank correlation and Wilcoxon signed-rank test were calculated using SPSS 11.5 software.Results (1)The SUV_(max) of T/WM,HIF-1α level and MVD in high-grade and low-grade tumors groups were 3.39±1.43,95.7% and 44.13±16.1 vs 1.46±0.55.55.6% and 18.83±7.07,respectively.The difierences of SUV_(max) of T/WM,HIF-1α level and MVD between two groups were statistically significant (t=-5.921,z=-3.938,t=-6.745,all P＜0.05).(2)Among 41 gliomas,the strong positive expression of HIF-1α was observed in 8,mederate in 9,weak in 15 and negative expression was found in 9,SUV_(max) of T/WM and MVD increased with increasing HIF-1α level.The differences of SUV_(max) of T/WM and MVD among 4 different groups were statistically significant (F=7.41,P＜0.05).(3) The MVD of all gliomas was ranged from 9.76 to 94.52,which correlated with SUV_(max) of T/WM(r=0.759,P＜0.05).Conclusions The SUV_(max) of T/WM correlates with HIF-1α level and MVD in gliomas.Therefore,~(18)F-FDG PET provides preoperatively a noninvasive assessment of hypoxia or angiogenesis in human glionma.

OBJECTIVETo evaluate in vitro effect of abnormal savda munziq (ASMq) on the proliferation and apoptosis of human hypertrophic scar fibroblasts (HSFs).

METHODSHSFs were divided into six groups to receive different treatments as group A (blank control group), group B-E (ASMq in different concentration), and group F(5-Fu). Each group contains six specimens. The HSFs were cultured in vitro. After culture for 48 hours, the CCK8 test and flow cytometry methods were used to detect the proliferation, cell cycle and apoptosis.

RESULTSThe proliferation of HSFs in the B, C, D and E groups was inhibited at G2/M period, while it was inhibited at G0/S period in group F (P < 0.05). The inhibition effect of ASMq (0.1-1.0 mg/ml) on the fibroblasts enhanced in a concentration-dependent manner. Flow cytometry analysis with annexin V-FITC and PI staining confirmed the apoptotic. When HSFs were exposed to ASMq at 1.0 mg/ml (group E) for 48 h, the percentage of apoptotic cells increased to (43.7 +/- 2.58)%, which was significantly higher than that of blank control group (2.2 +/- 0.59)%. The induced apoptosis effect was also increased in a concentration-dependent manner.

CONCLUSIONASMq has a inhibitory effect on the proliferation and an enhancement effect on the apoptosis of fibroblast. ASMq could be used as an effective drug for treatment of hypertrophic scar.

Apoptosis ; Cell Cycle ; drug effects ; physiology ; Cell Division ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cicatrix, Hypertrophic ; pathology ; Fibroblasts ; cytology ; drug effects ; Flow Cytometry ; Humans ; In Vitro Techniques ; Medicine, East Asian Traditional

OBJECTIVETo explore the therapeutic basis of high dose intravenous immunoglobulin (IVIg) in treatment of peripheral neuropathy induced by Campylobacter jejuni lipopolysaccharide (CJ LPS).

METHOD(1) IVIg (400 mg/kg x d) was given to the rats on the different days respectively during the immunization with CJ LPS. Histological study of sciatic nerve was performed on the 35 th day after immunization. The titer of anti-CJ LPS antibody in sera of immunized rats was measured by ELISA; IgG deposition was detected by immunohistochemistry and expression of TNF-alpha mRNA in the pathological nerves by in situ hybridization histochemistry. (2) When PBMCs were stimulated by CJ LPS in vitro, IVIg was added into culture medium at the doses of 1, 2.5, 5, and 10 mg/ml, respectively. Pathological examination of sciatic nerve was performed on the 7th day after perineural injection of the supernatants. Expression of TNF-alpha mRNA in PBMCs stimulated by CJ LPS in medium was detected by in situ hybridization histochemistry after adding IVIg.

RESULTS(1) The rate of abnormal fibers appearance in IVIg group (1.0%) was much lower than that of the control group (15.0%) after immunization with CJ LPS, P < 0.01. The titer of antibody in control group was 9 times higher than that of IVIg group. There was no expression of immunoglobulin and TNF-alphamRNA in peripheral nerves in IVIg group, but high expression was found in control group in which no IVIg was injected. (2) The expression rates of TNF-alphamRNA on the PBMCs in IVIg group (1.0%) was much lower than that of control group (9.5%). (3) When the PBMCs of normal rats were stimulated by CJ LPS, the expression rates of TNF-alphamRNA in PBMCs of 5 mg/ml IVIg group (3.0%) or 10 mg/ml IVIg group (2.0%) were much lower than that of 1 mg/ml IVIg group (15.0%) or 2.5 mg/ml IVIg group (11.5%), P < 0.01. The rate of abnormal fibers appearance in 5 mg/ml IVIg group (9.8%) or 10 mg/ml IVIg group (8.5%) was much lower than that of 1 mg/ml IVIg group (50.0%), 2.5 mg/ml IVIg group (41.0%) or control group (50.8%) after the perineural injection with the supernatants, respectively, P < 0.01.

CONCLUSIONThe therapeutic effect of high dose IVIg might be associated with inhibition of the humoral and cellular immunity simultaneously in peripheral neuropathy induced by CJ LPS.

Animals ; Enzyme-Linked Immunosorbent Assay ; Immunoglobulin G ; blood ; Immunoglobulins, Intravenous ; administration & dosage ; Immunohistochemistry ; In Situ Hybridization ; Leukocytes, Mononuclear ; drug effects ; metabolism ; Peripheral Nervous System Diseases ; genetics ; immunology ; therapy ; Rats ; Rats, Wistar ; Tumor Necrosis Factor-alpha ; analysis ; genetics

BACKGROUNDLaparoendoscopic single-site surgery (LESS) approaches have been reported for treating various kidney and pelvic procedures, and are feasible and effective in selected patients. In this study, we aimed to present the initial experience and evaluate the efficacy of laparoscopic radical prostatectomy performed through a single incision using a multichannel port.

METHODSBetween July 2010 and April 2011, six patients diagnosed with early stage prostate cancer underwent LESS radical prostatectomy (RP) in our institute. A multichannel port was inserted transperitoneally through a 2-cm umbilical incision. Specially articulating and flexible laparoscopic were used. Some technical tricks and points were applied during the operation to overcome the drawbacks and reduce the difficulties of this approach. Two continuous urethrovesical sutures in both sides were performed to complete both lateral aspects of anastomosis. The two ends of the suture threads were fixed by double Lapro-Clips, instead of the difficult knot-tying.

RESULTSTotal operative time was (265 ± 43) minutes. Mean blood loss was (230 ± 65) ml. All cases were completed successfully, without conversion to open surgery or adding additional abdomen ports. No patient required a blood transfusion and no intraoperative complications occurred. The Foley catheter was removed at the 14th day (range 12th - 16th) after surgery. At the 12th week of follow-up, all patients had an undetectable prostate-specific antigen level. Two patients used 2 or 1 pad for continence daily; other patients had achieved good continence.

CONCLUSIONIn selected cases, LESS-RP is feasible and effective; these technic points and the flexible-articulating instruments are helpful to reduce the operation difficulties.

Aged ; Humans ; Laparoscopy ; methods ; Male ; Prostatectomy ; methods

OBJECTIVETo investigate calcium, iron and magnesium intakes of preterm infants' mothers before and during pregnancy and calcium, iron and magnesium levels of preterm infants and their mothers in order to provide basis for studying the effect of nutritional factors on the occurrence of prematurity.

METHODSTwo hundred and forty matched cases (preterm infants and their mothers) and controls (term infants and their mothers) were recruited. A nutritional survey of calcium, iron and magnesium intakes was performed in the mothers before and during pregnancy. Calcium, iron and magnesium levels in maternal plasma and in cord blood, placenta, breast milk, meconium, and amniotic fluid were measured with axial view inductively coupled plasma optical emission spectrometry (ICP-OES).

RESULTSIron and magnesium intakes in preterm infants' mothers were significantly less than those in term infants' mothers before pregnancy (P<0.05). Iron and calcium intakes in preterm infants' mothers were also significantly less than those in term infants' mothers during pregnancy (P<0.05). Multivariate analysis of variance showed that iron and calcium levels of preterm infants' mothers were significantly lower than those of term infants' mothers (P<0.05). The preterm infants showed significantly lower iron and magnesium levels than term infants (P<0.05). Plasma levels of calcium, iron and magnesium in infants were positively correlated to maternal plasma levels of calcium, iron and magnesium (r=0.517, 0.622, 0.518, respectively; P<0.05).

CONCLUSIONSThe iron and calcium levels of preterm infants' mothers were lower than those of term infants' mothers, and the iron and magnesium levels of preterm infants were lower than those of term infants. The exact relationship between calcium, iron and magnesium levels and intakes before and during pregnancy needs to be explored further.

Calcium ; blood ; Calcium, Dietary ; administration & dosage ; Female ; Gestational Age ; Humans ; Infant, Newborn ; Infant, Premature ; blood ; Iron ; blood ; Iron, Dietary ; administration & dosage ; Magnesium ; administration & dosage ; blood ; Pregnancy ; blood

OBJECTIVETo evaluate the value of (18)F-FDG and (11)C-MET PET-CT scan in differentiation of brain ringlike-enhanced lesions on MRI imaging.

METHODSForty-one brain ringlike-enhanced lesions on MRI imaging including 30 brain tumors and 11 non-neoplastic lesions confirmed pathologically or clinically underwent (18)F-FDG and (11)C-MET PET-CT brain scan. Among them, 15 patients who were suspected to have brain metastasis received body scan by (18)F-FDG PET-CT. Both images were analyzed visually and semi-quantitatively.

RESULTSVisual analysis: for brain tumors the diagnostic sensitivity, specificity and accuracy of (18)F-FDG PET-CT was 53.3%, 72.7%, 58.5%, versus 96.7%, 90.9%, 95.1% of (11)C-MET PET-CT, respectively. All the primary foci in 9 patients with brain metastases were detected by body (18)F-FDG PET-CT scan. Semiquantitative analysis: There was a significant difference in the uptake between highly differentiated malignant and poorly differentiated tumors as well as non-neoplastic lesions for both tracers (P < 0.01), while between low-grade malignant tumors and non-neoplasm lesions, there was a difference in uptake only by (11)C-MET (P < 0.01). No significant difference between the uptakes in brain metastasis and glioblastomas was found by both tracers (P > 0.05).

CONCLUSIONBoth (18)F-FDG and (11)C-MET PET-CT are useful in differentiation of brain ringlike-enhanced lesions on MRI imaging. (11)C-MET PET-CT is more helpful than (18)F-FDG PET-CT in differential diagnosis of low-grade neoplastic from non-neoplastic lesions. Combination of (18)F-FDG and (11)C-MET PET-CT scans can improve the accuracy of differential diagnosis for brain ringlike-enhanced lesions on MRI imaging.

Acetates ; Adolescent ; Adult ; Aged ; Brain Abscess ; diagnosis ; Brain Neoplasms ; diagnosis ; pathology ; secondary ; Carbon ; Carbon Radioisotopes ; Child ; Craniopharyngioma ; diagnosis ; pathology ; secondary ; Diagnosis, Differential ; Female ; Fluorodeoxyglucose F18 ; Glioblastoma ; diagnosis ; pathology ; secondary ; Humans ; Lung Neoplasms ; diagnosis ; pathology ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Pituitary Neoplasms ; diagnosis ; pathology ; secondary ; Positron-Emission Tomography ; methods ; Radiopharmaceuticals ; Sensitivity and Specificity ; Young Adult

OBJECTIVETo investigate the effect of rat Schwann cell secretion on the proliferation and differentiation of human embryonic neural stem cells (NSCs).

METHODSThe samples were divided into three groups. In Group One, NSCs were cultured in DMED/F12 in which Schwann cells had grown for one day. In Group Two, NSCs and Schwann cells were co-cultured. In Group Three, NSCs were cultured in DMEM/F12. The morphology of NSCs was checked and beta-tubulin, GalC, hoechst 33342 and GFAP labellings were detected.

RESULTSIn Group One, all neural spheres were attached to the bottom and differentiated. The majority of them were beta-tubulin positive while a few of cells were GFAP or GalC positive. In Group Two, neural spheres remained undifferentiated and their proliferation was inhibited in places where Schwann cells were robust. In places where there were few Schwann cells, NSCs performed in a similar manner as in Group One. In Group Three, the cell growth state deteriorated day after day. On the 7th day, most NSCs died.

CONCLUSIONThe secretion of rat Schwann cells has a growth supportive and differentiation-inducing effect on human NSCs.

Animals ; Brain ; cytology ; embryology ; Cell Differentiation ; drug effects ; Cell Division ; drug effects ; Coculture Techniques ; Humans ; Rats ; Rats, Sprague-Dawley ; Schwann Cells ; secretion ; Sciatic Nerve ; cytology ; Stem Cells ; cytology

OBJECTIVETo explore the pathogenesis of the damage to peripheral nerves induced by Campylobacter jejuni exotoxin (CJT).

METHODS(1) Animal models: (1) The CJT was extracted from PEN 19-CJ and injected perineurally and intravenously to Wistar rats. (2) The sera and the supernatants of peripheral blood mononuclear cells (PBMCs), taken from the rats immunized with the CJT, were injected perineurally at sciatic nerves of experimental rats and intravenously, respectively. (2) Histopathologic study of sciatic nerves: the animals were sacrificed and their sciatic nerves were examined for tease fibers, transverse section with toluidine blues staining and electron microscopy. (3) Immunohistochemistry: sections of sciatic nerves of either normal rats or human which were incubated with CJT and the sciatic nerves with pathological changes induced by CJT were obtained for observation of the binding capability of CJT with peripheral nerves by SABC and FITC-immunofluorescence methods, and nucleic acid hybridization techniques for detection of TNF-alpha mRNA expression in pathological sciatic nerves samples.

RESULTS(1) Remarkable peripheral neuropathies with axon degeneration and/or demyelination were found in the nerves induced by both CJT injection perineurally and intravenously. The axon degeneration was more obvious. Pathological changes were identified in 76.8% (2,763/3,600) of teasing fibers after perineural injection, but only 9.6% (230/2,400) of fibers were damaged in control group (P < 0.01). The peak severity of fiber damage was found on the 3rd day after CJT intravenous injection with the incidence of abnormal fibers was 19.5% (390/2,000), and abnormalities of 15.5% (310/2000) on the 14th day. However, no abnormal changes were demonstrated in control group (P < 0.01). So was in the groups injected with anti-CJT sera and the supernatants of PBMCs compared with control (P > 0.05). (2) Binding of CJT to the nerve was found dominant in the sciatic nerves taken from normal rats or human either incubated with CJT or in the pathological sciatic nerves induced by CJT to various degrees. The binding of CJT to all these nerves was determined. (3) After intravenous injection with CJT, no histopathologic change could be found in the other viscera of the rats, with the exception of remarkable pathological change in peripheral nerves.

CONCLUSIONS(1) CJT could remarkably damage the peripheral nerves in rats. Specific pathogenicity of CJT to peripheral nerves was well shown, because no histopathologic abnormalities could be found in the other viscera, such as brain, liver and kidney etc. although there was remarkable pathological change along the peripheral nerve in the animals. (2) No immunological pathogenicity of CJT could be demonstrated in the nerves of rats after immunization with CJT.

Animals ; Antibodies, Anti-Idiotypic ; blood ; Bacterial Toxins ; immunology ; toxicity ; Campylobacter jejuni ; immunology ; Exotoxins ; immunology ; toxicity ; Gene Expression ; drug effects ; Peripheral Nerves ; drug effects ; metabolism ; pathology ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar ; Sciatic Nerve ; drug effects ; metabolism ; pathology ; Tumor Necrosis Factor-alpha ; genetics

OBJECTIVETo explore the DNA methylation levels of genome in cFb transdifferentiation induced by SiO2 in rats.

METHODSThe primary macrophages and fibrocytes of SD rats were co-cultured directly and indirectly, which were exposed to SiO2 at the doses of 25, 50 and 100 g/ml. The transdifferentiation of cFb was identified with immunohistochemical assay. The genomic DNA methylation levels of cFb were detected with HPLC.

RESULTSUnder the condition of indirect co-culture, as compared with control group, the genomic DNA methylation levels of cFb exposed to SiO2 at the doses of 25, 50 and 100 g/ml reduced by 19.9%, 26.9% and 30.3%, respectively (P < 0.05); as compared with cFb exposed to 100 g/ml SiO2, the genomic DNA methylation levels of cFb exposed to 5-aza-dC decreased by 22.0% (P < 0.05). Under the condition of ThinCert(TM) direct co-culture, as compared with control group, the genomic DNA methylation levels of cFb exposed to SiO2 at the doses of 25, 50 and 100 g/ml reduced by 22.2%, 30.2% and 36.7%, respectively (P < 0.05); as compared with cFb exposed to 100 g/ml SiO2, the genomic DNA methylation levels of cFb exposed to 5-aza-dC decreased by 20.6% (P < 0.05).

CONCLUSIONUnder the co-culture condition in vitro, SiO2 could reduce the genomic DNA methylation levels of cFb. The ThinCert(TM) direct co-culture can be used to study the silicosis fibrosis.

Animals ; Cell Transdifferentiation ; drug effects ; Cells, Cultured ; Coculture Techniques ; DNA Methylation ; Fibroblasts ; cytology ; drug effects ; Genome ; drug effects ; Lung ; cytology ; drug effects ; Male ; Rats ; Rats, Sprague-Dawley ; Silicon Dioxide ; adverse effects