AIMTo investigate the effects of protein tyrosine kinase on the inflammation and airway remodeling in lung of guinea pigs with bronchial asthma.

METHODS30 adult male guinea pigs were randomly divided into 3 groups (n=3): control group (C group), asthmatic group(A group)and genistein group (B group). Asthmatic model was established by ovalbumin intraperitoneal injection and ovalbumin inhalation. The total cell and the proportion of inflammatory cell in bronchial alveolar lavage fluid(BALF), inflammatory cell infiltration and index of remodeling of bronchiole were measured, respectively. The expression of p-tyrosine in lung tissue was examined by immunohistochemistry.

RESULTSThe total cell and proportion of eosinophil in BALF of A group were significantly higher than that of C group (P < 0.01), but compared with A group, the total cell and proportion of eosinophil in BALF of B group were much lower (P < 0.01). The number of eosinophile and lymphocyte of bronchiole in A group were significantly higher than that of C group (P < 0.01), but compared with A group, the number of eosinophile and lymphocyte in bronchiole of B group were much lower (P < 0.01). Compared with A group, the remodeling of bronchiole of B group was significantly relieved (P <0.01), there was no difference between B and C group (P > 0.05). Immunohistochemistry indicated that in A group the p-tyrosine was more positively expressed at the bronchial smooth muscle, bronchial epithelium, smooth muscle of vessel and inflammatory cell, especially at smooth muscle of bronchi and vessel and inflammatory cell than that of C group (P <0.01), there was no difference between B group and C group (P > 0.05).

CONCLUSIONPTK played a key role in inflammation and bronchial remodeling in lung of guinea pigs with bronchial asthma. The Protein tyrosine kinase inhibitor genistein could prevent and inhibit the inflammation and bronchial remodeling in lung of guinea pigs with bronchial asthma.

Airway Remodeling ; physiology ; Animals ; Asthma ; physiopathology ; prevention & control ; Genistein ; pharmacology ; Guinea Pigs ; Inflammation ; prevention & control ; Male ; Ovalbumin ; Protein-Tyrosine Kinases ; antagonists & inhibitors ; physiology ; Random Allocation

Objective To investigate the incidence and risk factors of surgical site infection(SSI)in neurosurgical patients in a tertiary first-class hospital,and provide reference for the prevention and control of SSI.Methods 47 neurological patients with SSI (49 patients developed SSI,2 were excluded from study due to the lack of appropriate control subject)from December 31 ,2011 to December 31 ,2012 were as infected group,and 94 patients without SSI (1 ∶2 matching)were as non-infected group,risk factors for SSI were analyzed retrospectively.Results There was no significant difference in general condition of two groups of patients (all P >0.05 );among 3 708 patients,49 (1 .32%)developed SSI;intracranial infection was the main type of SSI (89.80%);27 patients were performed ce-rebrospinal fluid (CSF)bacteriological detection,6 (22.22%)of whom were positive for CSF bacteriological detec-tion.Univariate conditional logistic regression analysis showed that risk factors for SSI in neurosurgical patients were operational risk assessment score (OR =2.04),frequency of preoperative antimicrobial use(OR =3.15 ),fre-quency of intraoperative antimicrobial use(OR=2.58),duration of operation(OR=2.70),surgical blood loss(OR=1 .72),indwelling drainage tube(OR=4.30),duration of indwelling drainage tube after operation(OR=2.06),and time for initial dressing change(OR=1 .66);Multivariate conditional logistic regression analysis showed that the in-dependent risk factors for SSI were frequency of preoperative antimicrobial use(P =0.03,OR =4.86),duration of operation(P =0.05,OR = 2.89 ),and time for initial dressing change after operation (P = 0.01 ,OR = 1 .92 ). Conclusion Risk factors for SSI in department of neurosurgery are multiple,duration of operation,duration of in-dwelling drainage tube after operation,and time for initial dressing change after operation are major risk factors.

Objective To investigate the correlation between setup error and couch position error in radiotherapy.Methods A total of 25 patients with thoracic and abdominal tumors who recently finished image-guided radiotherapy were randomly selected.The data on couch position during treatment were obtained through the record validation system, and then the couch position error was calculated.The Pearson correlation analysis was used to investigate the correlation between setup error and couch position error during treatment.Results In the ≥5 setup errors among the 25 patients, the correlation coefficient between random setup error and random couch position error was 0.83(P=0.00), and the correlation coefficient between systematic setup error and systematic couch position error was 0.36(P=0.11).Conclusions In radiotherapy, the random setup error is highly correlated with the random couch position error, while a moderate or low correlation exists between the systematic setup error and the systematic couch position error.

OBJECTIVETo investigate the dynamic expression and role of SENP1 (SUMO-specific proteases-1) in the pulmonary vascular wall of rat during the development of hypoxic pulmonary hypertension (HPH).

METHODSForty adult male Wistar rats were randomly divided into 5 groups (n = 8), and exposed to normoxia (Control group) or exposed to hypoxia for 3, 7, 14 or 21 d, respectively. The HPH models were established by normobaric intermittent hypoxia. Mean pulmonary arterial pressure (mPAP), right ventricle hypertrophy index (RVHI), and vessel morphometry were measured. Reverse transcriptase-polymerase chain reaction(RT-PCR) and in situ hybridization were used to determine the mRNA expression of SENP1. Immunohistochemistry and Western blot were used to determine the protein expression of SENP1.

RESULTSThe hypoxic rats developed pulmonary vascular remodeling in pulmonary arterioles after 7 d of hypoxia exposure. Pulmonary vascular remodeling in pulmonary arterioles significantly increased after 14 d of hypoxia. The level of mPAP in hypoxic rats increased significantly after 7 d of hypoxia, reached its peak after 14 d of hypoxic exposure. RVHI was markedly increased after 14 d of hypoxia. In situ hybridization and immunohistochemical analysis showed that SENP1 mRNA and protein were positively stained in control. SENP1 mRNA expression had little changes after exposure to hypoxia compared with the control, however, SENP1 protein expression was declined gradually after 7 d of hypoxia. The results of RT-PCR and Western blot showed that the same dynamic expression of SENP1 mRNA and protein in lung tissues of rats. Linear correlation analysis showed that SENP1 protein were negatively correlated with mPAP, pulmonary vascular remodeling index and RVHI.

CONCLUSIONUnder chronic hypoxia, SENP1 protein can be degradated. The dynamic expression of SENP1 protein may play a role in implicating in the development of HPH.

Animals ; Endopeptidases ; metabolism ; Hypertension, Pulmonary ; etiology ; metabolism ; Hypoxia ; complications ; metabolism ; Male ; Pulmonary Artery ; metabolism ; Rats ; Rats, Wistar

OBJECTIVETo explore the molecular mechanism of pain associated with chronic prostatitis and chronic pelvic pain syndrome (CP/CPPS) in the rat model of prostatic inflammation.

METHODSThirty-six male SD rats were equally randomized to an experimental and a control group, the former injected with 50 μl of 3% λ-carrageenan into the ventral prostate to make the model of non-bacterial prostatic inflammation, while the latter with the same volume of sterile saline solution. At 1, 2 and 4 weeks after modeling, the prostate, L6-S1 dorsal root ganglion (DRG) and spinal cord were harvested for examination of the expressions of the nerve growth factor (NGF), transient receptor potential ankyrin 1 (TRPA1), and calcitonin-gene-related peptide (CGRP) by immunohistochemistry and Western blot.

RESULTSThe expressions of NGF, TRPA1 and CGRP in the prostatic tissue were all significantly increased in the experimental group as compared with the control (P <0.05), with a gradual decrease with the prolonging of time (P <0.05). In the L6-S1 DRG and spinal cord, the expressions of NGF, TRPA1 and CGRP exhibited no significant differences between the experimental and control groups at 1 week after modeling (P >0.05) and kept at high levels in the experimental group at 2 and 4 weeks, though not significantly different from those at 1 week (P >0.05). Statistically significant differences were observed in the expressions of the three proteins in the experimental rats among different time points (P <0.05), but not between the two groups at any time point (P >0.05).

CONCLUSIONThe molecular mechanism of CP/CPPS can be evaluated in the rat model of prostatic inflammation established by injecting λ-carrageenan into the prostate. TRPA1 may play an important role in connecting the upstream and down-stream pathways of CP/CPPS-associated pain.

Animals ; Calcitonin Gene-Related Peptide ; metabolism ; Carrageenan ; Chronic Disease ; Chronic Pain ; metabolism ; Ganglia, Spinal ; metabolism ; Humans ; Male ; Nerve Growth Factor ; metabolism ; Pelvic Pain ; metabolism ; Prostatitis ; chemically induced ; metabolism ; Rats ; Rats, Sprague-Dawley ; Spinal Cord ; metabolism ; TRPA1 Cation Channel ; TRPC Cation Channels ; metabolism

OBJECTIVETo identify and separate the ventral root from dorsal root, which is the key for success of the artificial somatic-autonomic reflex pathway procedure for neurogenic bladder after spinal cord injury (SCI). Here we report the results of intra-operating room monitoring with 10 paralyzed patients.

METHODSTen male volunteers with complete suprasacral SCI underwent the artificial somatic-autonomic procedure under general anesthesia. Vastus medialis, tibialis anticus and gastrocnemius medialis of the left lower limb were monitored for electromyogram (EMG) activities resulted from L4, L5, and S1 stimulation respectively to differentiate the ventral root from dorsal root. A Laborie Urodynamics system was connected with a three channel urodynamic catheter inserted into the bladder. The L2 and L3 roots were stimulated separately while the intravesical pressure was monitored to evaluate the function of each root.

RESULTSThe thresholds of stimulation on ventral root were 0.02 ms duration, 0.2-0.4 mA, (mean 0.3 mA+/-0.07 mA), compared with 0.2-0.4 ms duration, 1.5-3 mA (mean 2.3 mA+/-0.5 mA) for dorsal root (P<0.01) to cause revoked potentials and EMG. Electrical stimulation on L4 roots resulted in the EMG being recorded mainly on vastus medialis, while stimulation on L5 or S1 roots caused electrical activities of tibialis anticus or gastrocnemius medialis respectively. The continuous stimulation for about 3-5 seconds on S2 or S3 ventral root (0.02 ms, 20 Hz, and 0.4 mA) could resulted in bladder detrusor contraction, but the strongest bladder contraction over 50 cm H2O was usually caused by stimulation on S3 ventral root in 7 of the 10 patients.

CONCLUSIONSIntra-operating room electrophysiological monitoring is of great help to identify and separate ventral root from dorsal root, and to select the appropriate sacral ventral root for best bladder reinnervation. Different parameters and thresholds on different roots are the most important factors to keep in mind to avoid damaging the roots and to assure the best results.

Adult ; Autonomic Pathways ; physiopathology ; Electric Stimulation Therapy ; methods ; Electromyography ; Electrophysiology ; methods ; Humans ; Male ; Muscle Contraction ; Muscle, Skeletal ; physiopathology ; Muscle, Smooth ; physiopathology ; Reflex ; Spinal Cord Injuries ; complications ; physiopathology ; Spinal Nerve Roots ; physiopathology ; surgery ; Thigh ; Urinary Bladder ; innervation ; physiopathology ; Urinary Bladder, Neurogenic ; etiology ; physiopathology ; surgery

OBJECTIVEA novel multiplex real-time RT-PCR kit was developed to detect EV71, CoxA16 and other human enteroviruses simultaneously with an internal amplification control to avoids false negatives, which used for hand, foot and mouth disease in the clinical diagnosis and epidemiological surveillance.

METHODSDesign specific primers and probes of EV71, CA16, other intestinal virus and internal amplification control, improve the extraction method of virus nucleic acid. Optimization the detection system of real-time quantitative PCR. Research the products of the accuracy, stability, precision, amplification efficiency and detection of linear range.

RESULTSThe primers and probes had high spicificity. The Viral RNA extraction effect of this Kit is as same as that of QIAamp Viral RNA mini Kit (QIAGEN company), but less reagent cost. The optimal concentrations of primers and probes are 0.2 micromol/L for all the upstream and downstream primers, 0.06 micromol/L for probes of other human enteroviruse, 0.08 micromol/L for probes of EV71 and CA16 respectively. The kit has good stability, accuracy and precision. The amplification efficiencies of EV71, CoxA16 and other human enteroviruses are 106% ,101% and 105% and the detection of linear range is from 10(9) copies/microl-10(2) copies/microl.

CONCLUSIONThe novel multiplex real-time RT-PCR kit for detecting EV71, CoxA16 and other human enteroviruses simultaneously with an internal amplification control has good stability, accuracy, precision and amplification efficiencies. So it has great value in clinical application.

Enterovirus ; isolation & purification ; Humans ; Reagent Kits, Diagnostic ; Real-Time Polymerase Chain Reaction ; methods

OBJECTIVETo study the effect of melatonin on proliferation and apoptosis of fibroblasts in human hypertrophic scar and its mechanism.

METHODSFibroblasts from human hypertrophic scar were isolated and cultured with DMEM medium containing 10% FBS, and then they were divided into control (C, added with ethanol), low concentration (LC, added with 1 × 10(-5) mmol/L melatonin), middle concentration (MC, added with 1 × 10(-3) mmol/L melatonin), and high concentration (HC, added with 1 mmol/L melatonin) groups according to the random number table. After being cultured for 24 hours, cell morphologic change was observed under microscope; XTT-PMS assay was used to examine cell proliferative activity; cell cycle and apoptosis were assessed with flow cytometry after double staining of FITC and PI, and the levels of cyclin E, p53, and Fas mRNA were determined with fluorescence quantitative RT-PCR. Data were processed with analysis of variance and LSD test.

RESULTS(1) Fibroblasts in C group were spindle-shaped with growth in colonies. Along with the increase in melatonin concentration, fibroblasts in LC, MC, and HC groups gradually dispersed, deformed and atrophied, with shrunk cellular membrane, and decrease in ratio of nucleus and cytoplasm. (2) Proliferative activity of fibroblasts in LC, MC, and HC groups decreased along with an increase in melatonin concentration (1.49 ± 0.15, 1.24 ± 0.20, and 0.92 ± 0.09), which were lower that in C group (1.79 ± 0.10, F = 67.61, P < 0.05). Cell ratios of S and G2/M phases in LC, MC, and HC groups decreased along with an increase in melatonin concentration, which were all lower than those in C group [(10.6 ± 1.1)%, (6.1 ± 1.2)%, (3.2 ± 0.8)% vs.(16.9 ± 1.3)%, F = 286.10, P < 0.05; (13.5 ± 1.1)%, (9.8 ± 1.0)%, (6.0 ± 0.7)% vs. (16.7 ± 1.6)%, F = 162.69, P < 0.05]. Apoptotic rates in early and late stages of LC, MC, and HC groups increased along with an increase in melatonin concentration, all higher than those in C group (with F value respectively 424.05, 236.44, P values all below 0.05). The expressions of cyclin E mRNA in LC, MC, and HC groups decreased along with an increase in melatonin concentration, which were lower than that in C group (1.58 ± 0.21, 0.90 ± 0.20, and 0.24 ± 0.12 vs. 2.90 ± 0.30, F = 266.79, P < 0.05), while the expressions of p53 mRNA and Fas mRNA showed opposite tendency (with F value respectively 10.11, 12.03, P values all below 0.05).

CONCLUSIONSMelatonin can inhibit proliferation and induce apoptosis of fibroblasts in hypertrophic scar through regulating the gene expressions of cyclin E, p53, and Fas.

Adult ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cicatrix, Hypertrophic ; metabolism ; pathology ; Cyclin E ; metabolism ; Female ; Fibroblasts ; drug effects ; metabolism ; pathology ; Humans ; Male ; Melatonin ; pharmacology ; Oncogene Proteins ; metabolism ; Tumor Suppressor Protein p53 ; metabolism ; fas Receptor ; metabolism

AIMTo investigate the effects of Nrf2 (Nuclear-E2 related factor) on gamma-glutamylcysteine synthase (gamma-GCS) in lung of guinea pigs with bronchial asthma.

METHODS20 adult male guinea pigs were randomly divided into two groups (n = 10): control group (C group) and asthmatic group (A group), asthmatic model was established by ovalbumin intraperitoneal and ovalbumin inhalation. The reactive oxygen piece (ROS), reduced glutathione (GSH), oxidant glutathione (GSSG) and total GSH in lung tissue were examined respectively. Inflammatory cell infiltration and index of remodeling of bronchiole were detected. In situ hybridization detected the gamma-GCS heavy subunit (gamma-GCS h) mRNA in lung tissue. Immunohistochemistry detected the expression of Nrf2 protein and gamma-GCS protein in lung tissue. RT-PCR measured the expression of Nrf2 mRNA in lung tissue. The activity of gamma-GCS was measured by coupled enzyme assay.

RESULTS(1) The number of eosinophils and lymphocytes in bronchiole of A group were significantly higher than that of C group (P < 0.05), the remodeling of bronchiole in A group was definite. (2) ROS (U/mg pro), GSSG (micromol/g pro) and total GSH in lung tissue of A group were significantly higher than that of C group (P < 0.01). The GSH/GSSG in lung tissue of A group was much lower than that of C group (P < 0.01), GSH in lung tissue showed no difference between A group and C group. (3) Immunohistochemistry indicated that Nrf2 protein and gamma-GCS protein were more positively expressed in A group than that in C group (P < 0.01). In situ hybridization discovered that the expression of gamma-GCS-h mRNA in lung tissue of A group was more positive than that of C group. (4) RT-PCR showed that the expression of Nrf2 mRNA was no difference between A group and C group (P > 0.05). (5) The activity of gamma-GCS of A group was (28 +/- 8)U which was significantly higher than that of C group (9 +/- 2)U (P < 0.01). (6) Linear correlation analysis indicated that in lung tissue of guinea pig with asthma there existed strongly positive relationship among ROS, GSSG and the expression of Nrf2, gamma-GCS mRNA, gamma-GCS protein, the activity of gamma-GCS, there existed strongly negative relationship among GSH/GSSG and the expression of Nrf2, gamma-GCS mRNA, gamma-GCS protein, the activity of gamma-GCS.

CONCLUSIONThere existed oxidative stress in lung of guinea pigs with bronchial asthma, which possibly positively regulated gamma-GCS via up regulating transcription factor Nrf2.

Animals ; Asthma ; metabolism ; Glutamate-Cysteine Ligase ; metabolism ; Guinea Pigs ; Lung ; metabolism ; Male ; NF-E2-Related Factor 2 ; metabolism ; Oxidative Stress ; RNA, Messenger ; genetics

OBJECTIVETo investigate the expressions of myogenic markers MyoD, myogenin,and desmin in skeletal muscle differentiation of human bone marrow mesenchymal stem cells (hBM-MSCs).

METHODSMyogenic markers MyoD, myogenin,and desmin of hBM-MSCs cultured in vitro were detected by immunofluorescence and RT-PCR. A total of 21 8-to-10 week-old immunosuppressed mdx mice were transplanted with 1x107 passage 5 of hBM-MSCs. The mice were euthanized 2-24 weeks after transplantation,and gastrocnemius muscle were analyzed for human MyoD, myogenin,desmin,and dystrophin (Dys) expressions by immunohistochemistry and RT-PCR.

RESULTSThe numbers of MyoD-,myogenin-,and desmin-positive cells per 100 hBM-MSCs were 23.5∓5.3, 30.7∓6.2, and 28.4∓5.7, respectively. MyoD, myogenin, and desmin mRNA was observed in passage 5 of hBM-MSCs. After two weeks of hBM-MSCs transplantation,a small number of MyoD-and myogenin-positive cells were observed in skeletal muscle of mdx mice,and desmin-positive cells were observed 4 weeks after transplantation. Expressions of MyoD and myogenin were detected in the muscle of mdx mice 2-4 weeks after hBM-MSCs transplantation, which reached a peak 12-16 weeks later. Desmin was expressed in the muscle of mdx mice 4-8 weeks after transplantation,with much more expression after 16 weeks of transplantation. A small number of Dys-positive cell and Dys mRNA expression were presented in the muscle of mdx mice 4 and 8 weeks after hBM-MSCs transplantation,respectively. The expression of Dys in the muscle of mdx mice increased gradually after transplantation.

CONCLUSIONhBM-MSCs have the potential of myogenic differentiation in vitro and contribute to myogenic conversion in xenogeneic animal,during which the up-regulation of MyoD and myogenin expressions may play an important role.

Animals ; Biomarkers ; Bone Marrow Cells ; cytology ; metabolism ; Cell Differentiation ; Cells, Cultured ; Desmin ; metabolism ; Humans ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Mice ; Mice, Inbred mdx ; Muscle, Skeletal ; cytology ; metabolism ; MyoD Protein ; metabolism ; Myogenin ; metabolism ; Up-Regulation