Objective To explore the diagnosis and treatment of clinical possible organizing pneumonia.Methods The medical records of 31 patients with clinical probable organizing pneumonia were retrospectively analyzed.The clinical presentation,radiographic results and treatment were collected and analyzed.Results Thirty-one patients with non-response to antibiotics were preliminary diagnosed as organizing pneumonia.By percutaneous lung biopsy or transbronchial lung biopsy the tuberculosis,fungi,suppurative inflammation,lung cancer and other diseases were ruled out in 24 patients,and 7 patients were diagnosed by perfect effect of corticosteroids treatment.Twenty-one patients had typical CT findings.All the patients responded rapidly and completely to the administration of corticosteroids.Six patients relapsed after the reduction or stop of corticosteroid in a follow-up time of 6-25 months.Conclusions Non-response to antibiotics,the typical imaging findings and lung biopsy ruling out other diseases are important for diagnosing organizing pneumonia.Rapid response to administration of corticosteroids may be helpful to the diagnosis.Decreasing the dose of corticosteroid too early may cause recurrence of organizing pneumonia.

Objective To explore the mechanism and effect of miR-146a on alveolar macrophages and to observe the changes of miR-146a expression in the LPS-induced alveolar macrophages. Method NR8383 alveolar macrophages were divided into LPS-stimulated group and control group, and the cells of former group were treated with LPS ( 1 μg/mL) and then incubated for 3 h, 6 h and 12 h, respectively. The level of TNF-α in the supernatant of cells was assayed by using enzyme-linked immunosorbent assay (ELISA), and the expression of miR-146a of cells was detected by using Real-Time PCR (TaqMan probe).Statistical analysis carried out by using SPSS 13.0 software package in which One-way ANOVA and Student's t-test were used. Results Compared with control group, the levels of TNF-α in the supernatant of cells were significantly increased 3 h, 6 h and 12 h after LPS challenge (P ＜ 0.01 ). The expression of miR-146a increased 6 h and 12 h after LPS stimulation in NR8383 cells( P ＜0.01 ), and it had an upward tendency.Conclusions The expression of miR-146a in alveolar macrophages increases after LPS-stimulation. It hints miR-146a may be involved in the regulation of the inflammatory responses produced by alveolar macrophages.

Objective To investigate the effect of Xing-Zhi-Yi-Nao (XZYN) particles on the expressions of Nogo and OMgp proteins in brain of rats after acute carbon monoxide (CO) poisoning.Methods A total of 120 Sprague-Dawley rats were randomly divided into normal group,CO poisoning group and XZYN particles treatment group (40 rats in each group).The rats in CO poisoning group and treatment group of acute CO poisoning were established by using an animal chamber,and then received hyperbaric oxygen therapy.Meanwhile,rats in treatment group were further given additional XZYN particles twice a day by gavage.At 1 day,1 week,1 month and 2 months after CO poisoning,the neurobehavioral score of rats was evaluated by a Morris water maze test and a shuttle box test,and the expressions of neurite outgrowth inhibitor (Nogo) and oligodendrocyte-myelin glycoprotein (OMgp) were investigated in rat brain tissue by immunohistochemistry staining and western blotting assay,respectively.Results Compared with those in normal control group((11.6±8.4)s,(41.8±4.4)%,(16.1±2.3)s,and (1.2±0.2)s),the escape latency in CO group was significantly prolonged ((14.1±6.1)s),and the T1/ T total was obviously decreased (23.6±2.4) %,the escape time ((26.3±3.8)s),the active escape latency ((2.3±0.3)s) were notably extended at 1 d (P<0.05).The cognitive dysfunction caused by CO poisoning was more obvious in the later stage of poisoning (from 1 week to 2 months,P<0.05).Compared with those in CO group,the escape latency was significantly shortened (from (3.5±0.6)s to (3.1±0.5)s),the T1/ T total was gradually increased (from (29.7±3.2)% to (36.7±3.2)%),the escape time (from (39.7±5.4)s to (18.1±2.0)s) and the active escape latency were obviously decreased (from (4.3±0.4)s to (2.1±0.2)s) in the later stage (>1 week) in Xing-Zhi-Yi-Nao treatment group (P<0.05).The expressions of Nogo and Omgp proteins in brain tissue in CO poisoning group were gradually increased as time went by.The increased expressions of Nogo and Omgp proteins were still observed at 1 month after CO poisoning.By contrast,XZYN particles could significantly improve cognitive function,reduce the expression of Nogo protein,and there was statistical difference compared with the poisoning group (P<0.05).However,the level of Omgp protein in XZYN treatment group was slightly lower than that in CO poisoning group,but there was no difference between the two groups (P>0.05).Conclusion The expression of Nogo and Omgp proteins may be associated with brain injury and demyelination in rats induced by CO poisoning.XZYN particles can down-regulate the expression of Nogo,and pave a way for the treatment of acute brain damage and delayed encephalopathy after CO poisoning.

Background and purpose:Carcinoma of the kidney is the most common malignant renal parenchymal carcinoma;its biological behavior is extremely complicated and is not sensitive to radiotherapy and chemotherapy.Studies have shown that the hypoxia-inducible factor HIF1-? and HIF2-? are related to the occurrence and development process of the clear-cell carcinoma of kidney.So,we intend to construct psilencer3.0-HIF-? siRNA recombinant plasmid in this study through RNA interference method,thereby providing an effective tool for further exploration of the role of HIF in the occurrence and development of clear-cell carcinoma of kidney.Methods:Design and chemically synthesize the DNA fragments of coding HIF-1? and HIF-2? siRNA and make them up into siRNA expression vector through gene recombination.Adopting the real-time quantitative PCR and Western blot tested the inhibitive effect of the constructed siRNA expression vector on mRNA and protein levels on the target gene expression.Results:After 786-0 cells transfecting with psilencer3.0-HIF-1?siRNA and psilencer3.0-HIF-2?,the inhibitory rate of mRNA expression of HIF-1? and HIF-2? reached 82.1% and 87.4% respectively,and OS-RC-2 cells transfecting psilencer3.0-HIF-1? siRNA and psilencer3.0-HIF-2?,the inhibition rate reached 91.2% and 81.2% respectively.The protein expression of the experimental group with 786-0 cells and OS-RC-2 cells transfecting HIF-1? interference plasmid and HIF-2? interference plasmid were lower at different levels than that from the blank control group.Conclusions:The constructed siRNA expression vector can effectively inhibit the expression of target gene HIF-1 and HIF-2 at mRNA and protein levels.

Objective To explore the regulation mechanism of X box binding protein 1 (XBP1)signal transduction pathway for TNF-α and effective approach in ischemia/reperfusion (I/R) injury of liver transplantation for short hairpin RNA (shRNA) interference used to gene therapy in liver graft.Methods Male Sprague-Dawley rats were divided into three groups: the cold ischemia transfection group (CIT), the in vivo transfection group (IVT) and the control group. Experiments of orthotopic liver transplantation were performed by two cuff method. The rats in CIT were perfused with XBP1-shRNA plasmid (pSIXBP1) during cold ischemia phase, those in IVT received the equivalent volume (2 ml) of pSIIRAK 4 after portal vein inoculation, and those in the control group were not subjected to any treatment. Rats were killed at 60 or 180 min after restoring reperfusion of hepatic portal vein.Histopathological damage degree of graft liver was observed by light microscope. The expression levels of XBP1 gene and protein were detected by RT-PCR and Western blotting. The activities of NF-κB and the serum TNF-α level were detected by ELISA. Results All the indexes including the degree of histopathological damage, the expression levels of XBP1 mRNA and protein and the TNF-α level were significantly decreased in CIT as compared with IVT and control group (P＜0. 05). However,there was no significant difference in NF-κB activity among the three groups (P＞0. 05). Conclusion The role of XBP1 pathway in TNF-α gene regulation and that of NF-κB pathway in rat liver I/R injury are two relatively independent aspects, and the depression of XBP1 expression with XBP1 shRNA through portal vein perfusion during cold ischemia phase could effectively alleviate graft hepatic I/R

Objective The expression and impaired function of ion channels might be one of the pathophysiological mecha -nisms responsible for diarrhea in inflammatory bowel disease ( IBD) .Proper animal model is the key to explore detailed pathophysiolog-ical process.The purpose of this study was to build a rat model of acute colitis induced by dextran sodium sulfate (DSS) in C57BL/6 mice and evaluate diarrhea-associated clinical , histological , pathological parameters and expressions of ion channel protein . Methods C57BL/6J mice of model group were treated with 4%DSS solution for 7 days to induce acute colitis.Mice body weight, stool moisture, stool consistency and the degree of hematochezia were recorded .The histopathological changes of mice colon specimens were observed visually and microcosmically, and the ion channel SLC26A3 protein was detected by Western Blot . Results All experimental mice survived.In the experiment, compared with control group , bloody diarrhea and weight lose occurred in model group , along with increased stool moisture ([73.30 ±8.31]% after experiment vs [44.32 ±6.42]% before experiment, P=0.004), and rapidly in-creased disease activity index (DAI) of acute colitis ([3.50 ±0.87] after experiment vs [1.0 ±0.00] before experiment, P=0.000).At the end of this experiment , compared with control group , the model group resulted in higher colonic damage score and pathological inflammation score (P=0.00, P=0.002), significantly shortened co-lon (P=0.00) and decreased expression of SLC26A3. Conclusion The intestinal mucosal injury and phenotypic features of 4%DSS-induced acute colitis are very similar to those of human ulcerative colitis .Impaired expression of intestinal ion transporter SLC26 A3 coexists with diarrhea in model group mice , and this model can support the research on mechanism of functional changes of ion channels in inflammatory diarrhea .

Objective To develop an ELISA method for determination of heparin-induced thrombocytopenia (HIT)antibody. Methods The compound formed between human platelet factor 4 (PF4)and heparin was used as the coating antigen,incu-bating the patients plasma with the coating antigen in the well,after washing,the second antibody labeled HRP was added in the well to incubate and washing again,the chromogenic substrates was added in the well to incubate,when the stop reaction was finished,the absorbance A450/A630 was detected,and the test results were judged according to standard,this method was compared with IBL method and was optimized and evaluated the performance.Results An indirect ELISA method was de-velop with the purified human PF4,the optimal dilution of sample and second antibody were 1∶100 and 1∶1 500 which de-tected by the orthogonal test,the intra-and inter-assay average coefficients of variation were 7.66% and 7.76%(<10%) respectively that detected by repeated measurement the three positive standard plasma.Through measureing the 100 healthy human plasm with no history of using heparin,the positive and negative predictive reference values were 0.304 and 0.456. IBL and this method detected 100 hemodialysis patients samples at the same time,and the result of statistical analysis was that,the sensitivity,speciality and accuracy of this method were 90%,97.78% and 90%,respectively.The negative and posi-tive predictive value were 81.8% and 98.88% respectively,and the difference was statistically significant [K=0.84(0.81~1)and Pexac=0.012<0.05].The difference was statistically significant,consistency was optimal,95% confidence interval was 92.59%~92.59%.Conclusion Comparing with the IBL,the method reported by this article had the similar perform-ance and good consistency,and it could satisfy the clinical detection and diagnosis of HIT patients.

OBJECTIVETo assess the effectiveness and safety of intratympanic versus systemic steroid therapy in the initial treatment of idiopathic sudden hearing loss.

METHODSAn extensive search of the literature was performed in Pubmed and other available database from January, 1980 to November, 2011. After filtering by the criteria of Cochrane Collaboration, a meta-analysis was conducted.

RESULTSNine studies met the criteria for meta-analysis, for idiopathic sudden hearing loss patients without diabetes received intratympanic steroid therapy, the improvement rate (RR = 1.11,95% CI = 0.96-1.28, P = 0.15) did not show any significance when compared with the patients received systemic therapy. While a significant difference of improvement rate occurred between intratympanic and systemic steroid therapy in the idiopathic sudden hearing loss patients with diabetes (RR = 1.24, 95% CI = 1.02-1.50, P = 0.03).

CONCLUSIONFor the initial therapy of idiopathic sudden hearing loss patients without diabetes, systemic steroid treatment still remains the first choice, but for the idiopathic sudden hearing loss patients with diabetes, intratympanic steroid treatment should be used for the initial treatment.

Administration, Oral ; Audiometry, Pure-Tone ; Dexamethasone ; administration & dosage ; therapeutic use ; Hearing Loss, Sensorineural ; drug therapy ; Hearing Loss, Sudden ; drug therapy ; Humans ; Methylprednisolone ; Steroids ; administration & dosage ; therapeutic use ; Treatment Outcome ; Tympanic Membrane

OBJECTIVETo investigate the effects of Fufang Banmao capusle on the proteome of SMMC-7721 cells and discover potential molecular mechanism of anti-cancer at molecular level.

METHODSMMC-7721 cells were treated by Fufang Banmao capusle serum prepared with serum pharmacological method; proteomic protocol involving 2-DE, image analysis and mass spectrometry were used to detect the proteins in cells influenced by Fufang Banmao capusle.

RESULTApproximately 450 protein spots in SMMC-7721 cells were resolved and detected in 2-D gel maps from pH3-10L IEF. 47 protein plots varied over 2-fold quantitively between treated sample and control sample were uncovered. 13 differentially expressed proteins spots were further identified by MALDI-TOF-MS analysis and four of them were successfully identified. Annexin A5, heatshock 70 x 10(3) protein 8 was significantly up-regulated in treated sample compared with control sample, while Eukaryotic translation initiation factor 5A and Peroxiredoxin-2 was significantly down-regulated in treated sample.

CONCLUSION4 differently expressed proteins associated with the proliferation, apoptosis, immunity of tumor were detected and they might provide clues for the coming research. The protocol of proteomics combined with serum pharmacological method is an effective platform to research complicated formulas in that it is capable of laying out many proteins associated with Fufang Banmao capusle.

Animals ; Annexin A5 ; metabolism ; Antineoplastic Agents ; isolation & purification ; pharmacology ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Coleoptera ; chemistry ; Drug Combinations ; Electrophoresis, Gel, Two-Dimensional ; Female ; HSP70 Heat-Shock Proteins ; metabolism ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Male ; Materia Medica ; isolation & purification ; pharmacology ; Peptide Initiation Factors ; metabolism ; Peroxiredoxins ; metabolism ; Proteome ; drug effects ; metabolism ; Proteomics ; methods ; RNA-Binding Proteins ; metabolism ; Rabbits ; Serum ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

OBJECTIVETo investigate the correlation of pathology, bone morphogentic protein (BMP) expression, CT value with the ossification of thoracic ligamentum flavum (TOLF) to afford the evidence to choose appropriate treatment methods.

METHODSTwenty-three patients aged 35 - 65 years old had TOLF in my hospital as case. Their courses of disease were 2 months to 9 years. The values of blood calcium, blood phosphorus and AKP in them were normal. The 5 peoples aged 21 - 35 years old who presented fracture of thoracic but not the ligamentum flavum ossification were selected as control. We excluded those who have DISH, ankylosing spondylitis, fluorosis and other disease related with TOLF. The lesion locus were scanned and mensurated by CT. The pathology characteristics were classified into immature ossification and mature ossification by general observation, histology examination. BMP were measured by the immunohistochemical (IHC) staining techniques.

RESULTSThe CT value was significantly higher in the case group (547.2 +/- 131.4) than controlled group (137.7 +/- 10.6) (t = 6.922, P = 0.000). Further, the CT value in the mature ossification (702.9 +/- 17.7) was significantly higher than the immature (480.5 +/- 180.2) (t = 5.623, P = 0.000). In addition, BMP both expressed negative in the mature ossification and the controlled group, but positive in the immature ossification. BMP expression was significantly different between the immature ossification and the mature (chi2 = 70.000, P = 0.000).

CONCLUSIONSThe CT values, pathological types and BMP expression results are similar to evaluate the ossification degrees of ligamentum flavum, and then could be indirectly judged the maturation degrees of TOLF by CT to confirm the treatment methods before operation.

Adult ; Bone Morphogenetic Proteins ; metabolism ; Case-Control Studies ; Female ; Humans ; Immunohistochemistry ; Ligamentum Flavum ; diagnostic imaging ; metabolism ; pathology ; Male ; Middle Aged ; Ossification, Heterotopic ; diagnostic imaging ; metabolism ; pathology ; Thoracic Vertebrae ; Tomography, X-Ray Computed ; Young Adult