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Objective To observe the effects of ischemic preconditioning of hindlimb (HIP) on the expressions of heat shock protein70 (HSP70) in spinal cord of rats after spinal cord injury (SCI). MethodSprague Dawleys rats were randomly( random number) divided into three groups (20 rats in each group).In sham injury group, rats received laminectomy without SCI; while in the SCI group, rats received traumatic SCI. In HIP group, rats received HIP 8 h before SCI. The SCI animal models were made by using modified Allen's weight-drop device (50 g·cm) on L3-L4 vertebrae. HIP was induced by compressing the two lower limbs of rats alternately with a tourniquet for three cycles of ten-minute ischaemia followed by ten-minute reperfusion. Rats were sacrificed 24 h and 48 h after injury separately. The protein and mRNA of HSP70 of involved spinal cord segments were determined by using RT-PCR, hybridization in situ and immunohistochemistry. Results There was weak expression of HSP70 mRNA in rats of sham injury group, and sporadic positive cells in spinal tissue were found by using immunohistochemistry. The expressions of HSP70 mRNA 24 h and48 hours after SCI in HIP group[(22.18±3.69) and (14.15 ±4.18)] were higher than those in SCI group[(13.97±4.46) and ( 8.73 ± 3.55 )], and the differences were statistically significant (F=16.06, P = 0.005 and F = 7.43, P = 0. 028). The protein levels of HSP70 24 h and 48 hours after SCI in HIP group [(16.71±4.02) and (9.85±2.20)] were higher than those in SCI group [( 14.85 ± 3.73)and (8.78 ±2.05)], and the differences were statistically significant (F =90. 13, P =0.032 and F=34.70, P = 0. 036). Conclusions HIP can increase the transcription and expression of HSP70 in spinal cord of rats following SCI and may have the protective effects on neural tissue.
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0.05),but these positive rates were notable decreased comparing with nPCR and immunohistochemistry techniques(P
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Objective To detect the pathological expressions of TNF-? and VEGF in interspinous ligament of patients with ankylosing spondylitis(AS) in active stage. Methods The expressions of TNF-? mRNA and VEGF mRNA in the interspinous ligament of patient with AS group and control group were detected by hybridization in situ. Their different results were compared by image analysis system and the correlation between TNF-? mRNA, VEGF mRNA and both AS disease active index (DAI) and C reactive protein (CRP) was analysed respectively. Results The expressions of TNF-? mRNA and VEGF mRNA in the interspinous ligament of patients with active AS were positive,but those of the control group were negtive. Statistical differences existed in the positive cell numbers and in image gray scale value of AS group (P
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Purpose:To study the significance of p53 protein, p53mRNA, and C erbB 2 protein expressions in Africa endemic and AIDS related Kaposi's sarcoma (KS).Methods:Immunohistochemistry and hybridization in situ were performed with p53 and C erbB 2 in 30 cases with endemic and AIDS related KS.The degree of pasitivity of p53 and C erbB 2 in KS tissues were measured with image analysis system. KS was classified into three types, i.e., angiomatous type, spindle cell type, and mixed type according to the proportion of blood vessel components and spindle cells in the lesions.Results:The degree of positivity of p53 protein was stronger in spindle cell type than that in angiomatous type ( P
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Objective To observe the survival,conversion and function preservation of mesenchymal stem cells(MSC) after transplantation.Methods MSCs from male Sprague-Dawley(SD) rats were isolated and cultured,then were transplanted to bone defect of female SD rats.Tissues were obtained at defferent periolds.Hybridization in situ was performed on the tissues using Y chromosome specific DNA probe labeled with digoxin to observe the transplanted cells on distribution,quantity and function.Results Allogeneic MSCs could survive and proliferate in the bone defect of recipients and formed bony callus with large quantity and good quality.After 90 days of graft,stem cells still survived and distributed in marrow and newly formed bone tissue.Conclusion After transplantation,allogeneic MSCs have a long-term surviving in marrow and new bone tissue and maintain the characteristic of ossification.
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Objective To investigate the expression of neuron nitric oxide synthase (nNOS) mRNA of brain tissue at early stage of intracerebral hemorrhage (ICH) in rats and the intervention effect of Didangtang.Methods 72 male Sprague-Dawley rats were randomly divided into four groups: intracerebral hemorrhage group, Didangtang treatment group, NOS inhibition group and normal saline group. Quantitate fresh autologous blood was infused into right caudate nucleus of rat sterotaxically to build up experimental ICH model and normal saline was instead in control group. The neurological function deficit scores were observed by Bederson method (3 grades) at 6 h, 24 h and 72 h after operation. At the same time points, the rats were sacrificed and the brain tissues were taken out for the measurement of nNOS mRNA by technique of hybridization in situ.Results Neurological function deficit scores of the rats were significantly improved both in intracerebral hemorrhage group and Didangtang treatment group at 72 h after operation(all P
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Objective To investigate the relationship between HPV infection and human colorectal carcinogenesis. Methods Colorectal carcinoma specimens from 72 Chinese patients were studied. DNA extracted from colorectal tissue was screened for HPV L1 by polymerase chain reaction (PCR), HPV subtype 6,11, 16, and 18 were detected by PCR using specific primers and in situ hybridization using specific probe. Results Twenty-four specimens out of 72 (33%) colorectal cancer were HPV L1 positive. The normal colorectal mucosa was HPV L1 negative. The location, infiltration and metastasis of colorectal carcinoma were all significantly related with HPV infection. The predominant HPV subtype was HPV 16,which was found in 58% of all HPV-positive colorectal carcinoma. Conclusion The presence of HPV DNA suggests that HPV may be involved in the carcinogenesis of colorectal carcinoma. HPV infection is closely related with the malignant potential of colorectal cancer.
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Objective: To investigate the effects of hexamethylene bisacetamide (HMBA) on apoptosis and the expression of bcl-2 in human mucoepidermoid carcinoma cell line MEC-1. Methods:MEC-1 cells were oxposed to 2 mmol/L HMBA or 1 mg/L 5 -Flourouracil (5 -FU ) for 72 hours. A poptosis index, the expression of bcl-2 protein and bcl-1 mRNA were determined by morphologic observation, terminal dUTP nick and labeling(TUNEL) stain, immunohistochemistry, in situ hybridization and image analysis. Results: Apoptosis indexes(%) in HMBA treated. 5-Fu treated and control cells were 68.5, 23.6 and 2.3 respectevely. The expression of bcl-2 protein and bcl-2 mRNA in HMBA treated cells was higher than in 5-Fu treated or the control cells(p