Objective To establish a model of injury to primarily cultured spinal cord neurons,mimicking the neuronal injury after complete transactional spinal cord trauma,for the sake of exploring changes in expression of an immediately-early gene,c-jun,in central nervous system injury. Methods Spinal cords were removed form fetal Wistar rats at the 14th gestation day and the neurons were cultured for 10 to 12 days.Then,mechanical injury was applied to the neurons by making regular scores on the culture disk under direct vision with the aid of a self-made standard template.Morphology of the injured neurons and changes in expression of c-Jun protein were observed before and at different intervals after injury. Results c-Jun expression was noted in neuronal nuclei 10 min after injury and its peak appeared at 2 hrs.Besides,the density of positive neurons bore evidently an inverse proportion with their distance from the scores.Conclusion\ Positive expression in injury neurons show that c-jun gene enters the nucleoli of injured neurons and takes the role of “the third messenger” at early time after neuronal injury.

Objective To clone injury-regeneration-related genes after primary neurons of spinal cord were injured in rat, and to elucidate the molecular mechanism of injury and regeneration in CNS. Methods An in vitro spinal cord primary neuron injury model was replicated in rat. The differentially expressed genes during injury and regeneration periods of spinal cord primary neurons were cloned by improved subtractive hybridization. Results Twenty-seven differentially expressed sequences were obtained, 23 were known genes and the others were unknown. Among these known sequences, cDNA for synuclein and clusterin, together with several other gene sequences, were closely related with nerve degenerative diseases. Conclusion Primary neuron injury-regeneration-related genes of the spinal cord can be successfully cloned by improved subtractive hybridization.

Objective To evaluate the STR profiling availability of formalin-fixed tissues and the detectable time limit of intact STR profile.Methods The different human tissues were fixed with 10-fold diluted commercial 40%formalin fixative for different duration under 15～20℃,and then DNA was extracted using the method of QIAamp~(R)DNA and IQ~(TM) DNA System.The extracted DNA was quantified with QuantifilerTM kit and amplified by both AmpFSTR identifiler kit and AmpFSTR MiniFiler kit.The STR profile was analyzed by GeneMapper ID v3.2 on 3100-Avant.Resuls The STR profiling availability of formalin-fixed tissues was relevant to the formalin fixing duration mainly.as well as the type of tissues and the template concentration and protocol of DNA extracting.The optimal ranges of template concentration is 1～3ng/μL and the QIAamp extracting method was preferable.There are differences in the degradation rate between various types of tissues in the unbuffered formalin fixative,and the lung tissue showed the slowest rate and liver and intestine tissues the fastest.Intact miniSTR profile of all the tissues detected could be obtained within 15 days duration of formalin fixing while intact STR profile could be obtained within 4 days.Conclusion The major factor that impact the availability of STR profiling of formalin-fixed tissues is the fixing duration in unbuffered formalin,as well as the type of tissues,method of extraction,concentration of PCR template and the kinds of STR loci.

To describe the clinical,radiological and pathological characteristics of idiopathic pulmonary alveolar proteinosis(I-PAP)and to evaluate the methods of diagnosis and treatment.Three patients were successfully diagnosed and treated in our hospital and the literature on the subject was reviewed.Three patients,two males and one female(mean age 46 years),were diagnosed averagely in 4 months.Two severe patients presented with progressive dyspnea and type I respiratory failure,and one mild patient only with dry cough and hypoxemia.Chest X-ray radiographs all showed perihilar "butterfly" shadow and chest CT scans showed diffused ground-glass opacities(GGO),typically with "map" changes and "crazy paving" patterns.All the patients underwent bronchoscope,branchoalveolar lavage fluid(BALF)had grossly opaque and/or milky appearance and its sediment was periodic acid-Schiff stain positive.Transbronchoscopic lung biopsy(TBLB)specimens were obtained and under light microscopy alveoli and some of the small bronchioles were filled with eosinophilic proteinaceous material with needle-like clefts.By electron microscopy numerous cellular debris and extracellular multilamellated bodies were found.Two severe patients were successfully treated with sequential whole-lung lavage and one required repeated lavages.I-PAP is rare and prone to be misdiagnosed.The radiological features may indicate the diagnosis and examinations of TBLB and BALF can make the accurate diagnosis.Whole-lung lavage is the most effective therapy by now and granulocyte-macrophage colony-stimulating factor(GM-CSF)may be beneficial in some patients.

To explore the clinical manifestation of lymphomatoid granulomatosis (LG) and the early diagnosis. Retrospective analyses of the clinical features of LG and pathology features by open lung bio-psies were conducted. The patient was a 42-year-old female with irregular fever,and her chest X-ray and computed tomography showed nodules with cavity and pleural effusion. Open lung biopsy proved LG. LG is seldom seen in clinic. Open lung biopsy is very important for pathology diagnosis. Early diagnosis and treatment are the key to improving survival in these patients. The therapeutic effect is good.

Objective To improve the diagnosis and treatment of pulmonary cryptococcosis.Methods The clinical data of 3 cases of pulmonary cryptococcosis were analyzed and reviewed.Results The cases were tested by percutaneous lung biopsy and were confirmed by histopathologic examination.The sputum cultures were negative and serum latex cryptococcal antigen agglutination tests were positive.Two of them had mild to moderate symptoms and were treated by fluconazol;the other with meningitis had severe symptoms and was treated by amphotericin B.All of them were clinically cured.Conclusion Percutaneous lung biopsy combined with latex cryptococcal antigen agglutination tests is helpful for diagnosis of pulmonary cryptococcosis.Patients with mild to moderate symptoms should be firstly treated by fluconazol and those with severe symptoms or meningitis,by amphotericin B.

BACKGROUND:During fracture healing, in addition to the need for appropriate biomechanical environment, the role of cytokines is also increasingly attracted attention.
OBJECTIVE:To study the effect of nerve growth factor and alpha-lipoic acid on fracture healing in rat models of femoral fracture.
METHODS:Sprague-Dawley rat models of femoral fracture were established. Seventy-two rats were randomly divided into three groups. In the control group, rats were intramuscularly injected with physiological saline. In the nerve growth factor group, rats were intramuscularly injected with nerve growth factor 200 ng/kg, once a day. In the combined therapy group, rats were intramuscularly injected with nerve growth factor 200 ng/kg and oral y taken alpha-lipoic acid 25 mg/kg, once a day. At 1, 2 and 3 weeks after administration, bony cal us volume was measured. Enzyme linked immunosorbent assay was used to measure serum bone morphogenetic protein-2 levels. Western blot assay was utilized to detect bone morphogenetic protein-2 protein expression at the broken end of fracture. Semi-quantitative RT-PCR was applied to examine vascular endothelial growth factor mRNA expression.
RESULTS AND CONCLUSION:(1) At 1 week after administration, no significant difference in bony cal us volume was detected among the three groups. Serum bone morphogenetic protein-2 level, bone morphogenetic protein-2 protein expression, and vascular endothelial growth factor mRNA expression were significantly higher in the nerve growth factor group and combined therapy group compared with the control group (P<0.05), but no significant difference was found between the two groups. (2) At 2 weeks after administration, the amount of cal us, serum bone morphogenetic protein-2 levels, bone morphogenetic protein-2 protein expression, and vascular endothelial growth factor mRNA levels were significantly higher in the nerve growth factor group and combined therapy group compared with the control group (P<0.05). Above expression levels were higher in the combined therapy group than in the nerve growth factor group (P<0.05). (3) At 3 weeks after administration, serum bone morphogenetic protein-2 levels, bone morphogenetic protein-2 protein expression, and vascular endothelial growth factor mRNA levels were significantly decreased in the nerve growth factor group. However, above expression levels were stil high in the combined therapy group, and significantly higher than in the nerve growth factor group (P<0.05). (4) These results indicate that nerve growth factor combined with alpha-lipoic acid had better effects on the fracture healing compared with the nerve growth factor alone.

AIM: To investigate the role of expression of peroxisome proliferator-activated receptor ?(PPAR ?) in pathogenesis of rat fatty liver.METHODS: The rats were treated with a low dose of carbon terachloride (CCl 4) and fed a high fat diet to produce fatty liver. We determined the concentrations of triglyceride (TG), total cholesterol (TC), free fatty acid (FFA) in liver and the alanine aminotransferase (ALT) activity, tumor necrosis factor-? (TNF-?), FFA in serum and the degree of hepatocytic steatosis. Total RNA of liver was extracted, and the expression of PPAR ? were analyzed by semi-quantitative RT-PCR method. RESULTS: In model group, the hepatocytic PPAR ? mRNA expression decreased to 0 41?0 28, compared to 1 41?0 29 in the control group ( P

Objectives To investigate the effect of different dosages of low molecular weight heparin on acute pulmonary embolism and inhibition of pulmonary intimal hyperplasia in immature rats. Methods 90 male immature SD rats were randomly divided into ifve groups: sham group, pulmonary embolism group, low-low molecular heparin group (L-LMH), medium-low molecular heparin group (M-LMH) and high-low molecular heparin group (H-LMH). The model of acute pulmonary embolism was established through jugular vein injection with gel-foam solution. The rates in the L-LMH, M-LMH, H-LMH groups were treated with low molecular weight heparin by subcutaneous injection after surgery with a dosage of 0 . 005 ml/kg, 0 . 01 ml/kg, 0 . 02 ml/kg, twice a day. Animals in the control group were given saline injection. Arterial blood gas, pulmonary artery pressure (mPAP), right ventricular pressure (RVP), wall area/tube area, wall thickness/tube diameter, and the expression of PDGF-B and MCP-1 at gene and protein levels in lung tissue were detected on the 7 th ( 7 d), 14 th ( 14 d) and 28 th ( 28 d) after opration. Results There were signiifcant differences of PaO 2 among 5 groups on 7 d, 14 d and 28 d. PaO 2 in group M-LMH ( 105 . 1 ± 4 . 6 mm Hg) were signiifcantly higher than that of embolization group, L-LMH, but not H-LMH group at 28 d. mPAP of M-LMH group was lower than that in the other three intervention groups, but showed no signiifcant difference compared with sham group (P?>0 . 05 ). There were signiifcant differences of RVP on 7 d and 14 d. PDGF-B, MCP-1 of M-LMH group were signiifcantly lower compared with the other three intervention groups (P?0 . 05 ). Wall area/tube area, wall thickness/tube diameter scores of M-LMH group had no signiifcance differences compared with sham group on 28 d (P?>?0 . 05 ). Conclusion Medium dose of low molecular weight heparin could ameliorate the acute pulmonary embolism and inhibit the proliferation of pulmonary arteries in rats.

To study the mechanisms whereby cisplatin suppresses survival of human esophageal squamous cell carcinoma cells.The cytotoxicity of cisplatin in cisplatin-resistant cell line EC109 /CDDP and its parental cell line EC109 was measured by cell viability assay.Western blotting was used to investigate the protein expression of to-tal p53 and phosphorylated p53 at Ser15.Colony formation assay was employed to evaluate the ability of cells to recover from treatments and form colonies.The results indicated that EC109 /CDDP cells were more resistant to cisplatin-induced cytotoxicity than EC109 cells,with the IC50 values of (20.4 ±4.4)μmol /L and (5.7 ±0.1 )μmol /L,respectively.Although cisplatin did not alter the total protein level of p53,it obviously increased the phosphorylation of p53 at Ser15.Cisplatin inhibited survival of both EC109 /CDDP and EC109.Notably,inhibition of p53 by Pifithrin-αsignificantly promoted recovery of cisplatin-treated EC109 and EC109 /CDDP cells to differ-ent degrees.In this respect,p53 protein was found to be activated in response to cisplatin treatment in both EC109 /CDDP and EC109,which may contribute to the cytotoxic effect of cisplatin.