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Objective@#To investigate the methylation status and expression of FOXP3 in CD4+ T cells of patients with chronic hepatitis B (CHB).@*Methods@#Peripheral blood mononuclear cells (PBMCs) from 59 CHB patients and 22 healthy controls (HC) were collected. The percentage of CD4+ CD25+ Foxp3+ Tregs in CD4+ T cells was estimated by flow cytometry. FOXP3 expression was measured by quantitative real time-polymerase chain reaction (RT-qPCR) and Western blotting. The methylation status of FOXP3 was determined by methylation-specific polymerase chain reaction.@*Results@#The percentage of CD4+ CD25+ pFoxp3+ Tregs in CD4+ T cells, FOXP3 mRNA and protein expression levels were significantly higher in patients with CHB than HCs (P<0.05). Meanwhile, the methylation frequency of FOXP3 was significantly lower in CHB patients than HCs (P<0.05). FOXP3 mRNA levels and the percentage of CD4+ CD25+ Foxp3+ Tregs were significantly lower (P<0.05) in patients with gene methylation than those without.@*Conclusions@#Aberrant demethylation of FOXP3 gene existed in CD4+ T cells of CHB, which contributed to an elevation in FOXP3 expression and percentage of CD4+ CD25+ Foxp3+ Tregs. It might provide a new target for prevention and treatment of CHB.
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Objective To evaluate the feasibility of needle-based confocal laser endomicroscopy (nCLE) for imaging of intra-abdominal tissues and organs in rabbit models in vivo. Methods The nCLE miniprobe was inserted through the 19-gauge needle into various intra-abdominal tissues and organs[omentum majus, liver, pancreas and psoas major (skeletal muscle)]. The nCLE images were acquired and real-time sequences of respective locations were recorded. Finally, nCLE image characteristics were compared with histopathologic findings. Results nCLE was successfully performed in intra-abdominal tissues and organs of five rabbit models. The microscopic structures of cells, glands and microvessels in the omentum majus, liver, pancreas and psoas major ( skeletal muscle) were visualized clearly, respectively. Characteristics of various intra-abdominal tissues and organs were displayed on nCLE images, which were correlated well with histological findings. Conclusion Imaging of intra-abdominal tissues and organs with nCLE in vivo is feasible in future clinical practice.
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Objective To observe the role of brain-derived neurotrophic factor (BDNF) in the plasticity of enteric nervous-smooth muscle system,and to investigate the effects of BDNF induced plasticity on gastrointestinal motility in mice.Methods Male hybrid BDNF knockout (BDNF+/) mice and wild type (BDNF+/+) mice were selected,eight in each group.Gastrointestinal motility of BDNF+/+ mice and BDNF+/ mice were tested and compared.Longitudinal muscle strips of mice colon smooth muscle were prepared.The effects of carbachol (1 × 10 5 mol/L) and BDNF (1 × 10 7 mol/L) on contractile function of muscle strips were observed.And the effects of tetrodotoxin (TTX,1 × 10-6 mol/L) on BDNF induced contractile function of muscle strips were also studied.The changes of the density of mice intestinal myenteric plexus and the expression of smooth muscle α-actin (α-SMA) in colon smooth muscle were detected by immunohistochemical techniqne.The ultrastruetural alterations of myenteric plexus,neuromuscular junction (NMJ) and smooth muscle cells were detected by transmission electron microscope (TEM).T-test or Rank sum test was performed for comparison between groups.Results Number of feces particles and water content in feces of BDNF+/-mice ((3.80±0.75) and (39.60±1.47)％) were both lower than those of BDNF+/+ mice ((6.30± 1.03) and (51.00± 1.61) ％),and the differences were statistically significant (t=4.792,12.827;both P＜0.05).Carbachol (1 × 10-5 mol/L) could significantly increase contraction activity of smooth muscle of BDNF+/+ mice (R =3.26 ± 0.43) and BDNF+/-mice (R=2.15 ± 0.36),and the difference was statistically significant (t=15.754,9.632;both P＜0.05).The effects on contraction exciting of smooth muscle strips of BDNF+/+ mice were more significant than BDNF+/ mice,and the difference was statistically significant (t =5.972,P＜0.05).BDNF could significantly increase contraction of muscle strips of BDNF+/+ mice and R value increased from 1 to 1.41±0.09,and the differences were statistically significant (t=13.674,P＜0.05).TTX could obviously inhibit the excitatory effects of BDNF,R value decreased from 1.41 ± 0.09 to 1.03 ± 0.04 (t=11.692,P＜0.05).The density of myenteric plexus of BDNF+/ mice (median 5.8％,interquartile range 4.2％-7.0％) was significantly lower than that of BDNF+/+ mice (median 9.0％,interquartile range 7.1％-10.8％),and the difference was statistically significant (Z =3.730,P＜ 0.05).The expression of α-SMA of BDNF+/-mice (median 33.4％,interquartile range 28.8％-38.5％) was significantly lower than that of BDNF+/+ mice (median 44.6％,interquartile range 39.2％-48.8％),and the difference was statistically significant (Z=4.565,P＜0.05).The results of TEM indicated ultrastructural alterations of myenteric plexus,NMJ and smooth muscle in BDNF+/-mice.Conelusionss BDNF could induce the plasticity of morphology and function in enteric nervous-smooth muscle system,which may play an important role in mice gastrointestinal motility.
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Objective To evaluate the quality of the examination paper of the theory of Clinical Diagnostics,to explore and reflect on the teaching methods,so as to improve the quality of teaching.Methods Finals results of Clinical Diagnostics including 40 clinical undergraduates of Capital Medical University were analyzed.SPSS 20.0 was used to make analysis of the frequency,means statistics and normality of the examination paper.Kuder Richardson/Cmnbacha formula,percentage unification methods and so on were used to calculate confidence,validity,difficulty and degrees of distinction.Acquisition of relevant knowledge was assessed according to score distribution,while test paper quality was evaluated based on indicators including confidence,validity,difficulty and degrees of distinction.Results Test scores of 40 students were between 61 to 96 (83.64 ± 8.07).The degree of confidence (γ) for choice questions and subjective questions was 0.65 and 0.59 respectively;The validity (V) was 0.27;The overall difficulty (P) of the examination was 0.84;The degrees of distinction (D) were between 0.16 to 0.30.And the total points losing rate was 16.36％.Conclusion The examination is of medium difficulty and good degree of distinction,but the teaching strategies still need further adjustment in order to improve the students' ability of flexible application of the basic knowledge.
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Objective To investigate the correlation between the expression changes of brain-derived neurotrophic factor (BDNF) in colon mucosa and abdominal pain in irritable bowel syndrome (IBS). The density of nerve fiber in colon mucosa and ultrastructural alterations of nerve fiber in IBS were also observed. Methods From September 2008 to January 2010,the IBS patients who visited the department of gastroenterology of our hospital and met the Rome Ⅲ diagnosis criteria were selected and divided into IBS with diarrhea (D-IBS) and IBS with constipation (C-IBS) according to their clinical features. The patients with colon polyps detected by colonoscopy in our hospital were selected as control group. All subjects were asked to fill in Self-Rating abdominal pain or abdominal uncomfortable Scale according to abdominal symptom in the last 2 weeks before visit and underwent colonoscopy. Four biopsy specimens were taken from the colon mucosa of rectosigmoid junction. Ofwhich,two specimens were for protein isolation and detection of BDNF expression level,one specimen was used for PGP 9. 5 immunohistochemistry staining in paraffin slices. Another specimen was used to observe the ultrastructure changes of nerve fiber in colon mucosa under transmission electron microscopy. Results Total 40 IBS patients were enrolled in this study,of those 21 were D-IBS patients,19 were C-IBS patients,and 21 were controls. The abdominal pain severity score and frequency score of IBS patients were (2. 3±0. 8) and (2. 1±0. 7),which were significantly higher than those of control group (0. 4±0. 7 and 0. 3±0. 5,P＜0. 001). Compared with the control group,the BDNF expression in colon mucosa was significantly elevated in IBS patients (P= 0. 003 ),and which correlated with the severity and frequency of abdominal pain/discomfort (r=0. 57,P＜0. 001and r=0. 46,P= 0. 003,respectively). The immunohistochemistry result indicated that the nerve fiber density in colon mucosa of IBS patients was significantly higher than that of controls,and there were ultrastructural changes of colon mucosal nerve fibers in IBS patients. Conclusion Increased colon mucosal BDNF expression may be associated with abdominal pain symptom in IBS patients. The impaired ultrastructural of mucosal nerve fibers may cause the increased BDNF expression in colon mucosa,and result in the increased mucosal nerve fiber density in IBS patients.
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BACKGROUND: Osteoporosis is one of the complications of spinal cord injury, but its mechanism is unclear, different scholars have different points about whether all bones above and below the level of trauma are affected after spinal cord injury (SCI).OBJECTIVE: To observe the ultrastructure of bone tissue of secondary osteoporosis after spinal cord injury and the change of serum biochemical indexes, to analyze the suffered condition and injured degree of the bones above and below the level of the trauma.DESIGN: Randomized controlled experimentSETTING: Laboratory of Animals, and Department of Laboratory Medicine of the Second Clinical Hospital of Jilin University; Transmission Electron Microscope Center, Norman Bethune Division of Medical Sciences of Jilin UniversityMATERIALS: This experiment was conducted at the Laboratory of Animals, Second Clinical Hospital of Jilin University from May 2002 to May 2003. Totally 110 male Wistar rats, aged 4 to 5 month-old, with the body mass of (300 -320)g were involved.METHODS: Ten rats were randomly chosen from 110 male Wistar rats and were set as 0 week blank control group, the ther rats were randomly divided into experimental group and control group, there were 10 rats in week 1, 2, 3, 7 and 11 control group and experimental group separately.1 mL/L of pentobarbital natrium was intraperitoneal injected to perform anesthesia, in the control group, lamina with dura intact of rats were removed only, without spinal cord injury at the level of tenth thoracic vertebrae. In the experimental group, lamina of rats was also removed, the spinal cord injury model of the rats were made by the method of Allen's.4 mL venous blood was collected from the animals at the end of postoperative week 0, week 1, week 2, week 3, week 7 and week 11, then the animals were put to death, perform blood centrifuging .We observe the change of serum concentration of calcium, phosphorus and alkaline phophatase (ALP) with 7170A HITACHI auto-biochemistry analyzer. Right side humerus, tibia in week 0 blank control group, week 7 experimental group and week 11 experimental group were chosen, then 25 mL/L ethylene dinitrilotetraacetic acid was used for decalcium for 1 month for preparingthe sections. We observe the change of ultrastructure of osteocytes at the sites of tibial plateau and the surgical neck of the humerus after stained by uranyl acetate and lead citrate under transmission electron microscope.biochemistry detection: the level of phosphorus at week 2 of the experiment was significantly lower in the control group than experiment group [(1.54±0.21),(2.76±0.16)mmol/L, (P ＜ 0.01)]; the level of calcium at week 3 in the experiment group was significantly higher than in the control group [(2.52±0.06),(2.35±0.12)mmol/L, (P ＜ 0.01)];the level of alkaline phophatase at week 7 in the experiment group was significantly higher than in the control group [(155.86±20.42), (129.25±7.30)Nμ/mg,operative week 7, the tibia sample showed that the osteocyte separated from the osseous lacuna, the nucleus showed anomaly, fluffed materials appeared. Mitochondrion swelled, rough endoplasmic hollowed; At week 11, osteocyte separated from the osseous lacuna. At postoperative 7 week,the humerus sample showed that osteocyte separated from osseous lacuna,some nucleus pycnosis and rough endoplasmic hollowed; At postoperative week 11, the above changes still were seen, but the degree of cellular swelling and hollowing were lighten.CONCLUSION: At the early stage of spinal cord injury, the activity of osteoclast is increased and the activity of osteoblast is decreased, which leads to the increase of bone absorption, the bone formation is largely weakened. The bones above and below the level of trauma are both affected after spinal cord injury, but different extents of osteoporosis can be seen in different bones .The change of ultrastructure of osteocytes is remarkable when osteoporosis has happened.