Objective To study the neuroprotective effects of neuregulin-1?(NRG-1?) on the nervous behavioral function,cerebral infarction volume,brain water content(BWC),neuronal apoptosis and aquaporin-4(AQP-4) expression in astrocytes after cerebral ischemic reperfusion and the related mechanism in mice.Methods Intraluminal thread methods were applied to establish the middle cerebral artery occlusion reperfusion models in the mice.Neuregulin-1?(2?g / kg) was injected into the internal carotid artery for treatment.The nervous behavioral function was evaluated with Bederson's test.The cerebral infarction volume was observed with tetrazolium chloride staining.The BWC was measured by dry-wet weight comparing.The apoptosis positive cells were counted by immunofluorescence assay.The expression of AQP-4 was determined by immunohistochemical assay.Results Nervous behavioral malfunction appeared in all the mice with left middle cerebral artery occlusion and/or reperfusion.The infarction focus showed in the ischemic hemisphere after the injury.The BWC,the number of neuronal apoptosis cells and AQP-4 expression in astrocytes were higher than those in the sham group. In the NRG-1? treatment group,the nervous behavioral function was improved 24 hours after ischemia,the number of apoptosis positive cells reduced and the infarction volume decreased significantly compared with the control group(P0.05).In the groups of reperfusion for 22,46 and 70 hours,the five indexes mentioned above were significantly different from those in the corresponding control groups(P

BACKGROUND: Most animal experiments of transgene are derived from mice; therefore, it is necessary to establish a focal cerebral ischemiareperfusion model and significant to prevent and cure ischemic cerebrovascular diseases.OBJECTIVE: To establish a convenient and reliable model with middle cerebral artery occlusion reperfusion (MCAO/R) in mice.DESIGN: Randomized controlled animal study.SETTING: Institute of Cerebrovascular disease, Affiliated Hospital of Medical College of Qingdao University.MATERIALS: Twenty healthy BALB/c mice, of both genders, weighing 25-30 g, of SPF grade, were divided into sham operation group (n=5), ischemia group (n=10) and 22-hour reperfusion group (n=5) on the basis of digital table. In addition, according to digital table, 130 healthy male Kunming mice were divided into sham operation group (n=10), 24-hour ischemia group (n=30), 2-hour ischemia/22-hour, 46-hour and 70-hour reperfusion groups with 30 in each group; meanwhile, 30 female mice were divided into sham operation group, 24-hour ischemia group and 2-hour ischemia/22-hour reperfusion group with 10 in each group. All Kunming mice were weighing 25-30 g and of SPF grade.METHODS: The experiment was carried out in the Institute of Cerebrovascular Disease, Affiliated Hospital of Medical College of Qingdao University from July 2005 to March 2006. The 6-0 suture with silica gel covered at an end was sent from the left external carotid artery (ECA) into internal carotid artery (ICA) till arriving at the initiation of middle cerebral artery (MCA) to block the blood stream in it, then drawing the suture from ICA 2 hours after occlusion to accomplish reperfusion. Mice were cut off their heads in sham operation group at 24 hours after operation, in ischemia group at 24 hours after blocking blood stream and in reperfusion group at 24, 48 and 72 hours after operation. Reliability of models was evaluated with neurology score and tetrazolium chloride stain. Longa standard scores: neurology score ≥ 1 point was regarded as successful models;coronal sections of brain tissue were stained with tetrazolium chloride, and the white region was regarded as infarcted volume.MAIN OUTCOME MEASURES: Neurology score and infarcted volume after staining of triphenyltetrazolium chloride in brain tissue.RESULTS: All mice were involved in the final analysis. ① Successful rate was 20% of BALB/c mice, 66.7%-73.3% of male Kunming mice and 40%-50% of female Kunming mice. ② Brain sections of BALB/c mice in sham operation group were orange at both sides of cortex and infarction focus was not observed. A big infarcted volume was observed on brain sections of mice in ischemia group, and infarcted volume counted for 50%-70% as homonymy hemisphere on optochiasmatic coronal sections. The condition of Kunming mice was similar to that of BALB/c mice, but infarcted volume counted for 40%-65%. In addition, condition in ischemiareperfusion group was similar to that in ischemia group. A big infarcted volume was observed on brain sections, and infarcted volume counted for 50%-75% as homonymy hemisphere on optochiasmatic coronal sections.The condition of Kunming mice was similar to that of BALB/c mice, but infarcted volume counted for 40%-65%.CONCLUSION: The model with MCAO/R in mice characterizes by relatively smaller trauma, and the ischemic region is stable; therefore, it can be used to accurate timing control of ischemia/reperfusion. This model is an ideal one for researching pathophysiological changes, prognosis and therapy in cerebrovascular disease.

Objective To investigate the probable risk factors for type 2 diabetic patients complicated nonalcoholic fatty liver disease (NFLD)in elderly, through comparing the body composition, serum lipid profile, incidences of abdominal obesity and metabolic syndrome (MS) between elderly type 2 diabetic patients with and without NFLD. Methods The enrolled elderly type 2 diabetic patients were divided into NFLD group (n=83) and non-NFLD group (n=85). Their clinical data including body composition, serum lipid profile, incidences of abdominal obesity and MS were analyzed retrospectively and compared. Results Compared with non-NFLD group, the BMI [(26.9±2.5) kg/m~2 vs. (24.1±2.5) kg/m~2, P=0.000], waist-hip ratios (WHR) ((0.92±0.07) vs. (0.87±0.06), P=0.000], total body fat percentage [(29.6%±6.6%) vs. (25.3%±5.5%),P=0.000], abdominal fat [(11.0±2.5) kg vs. (8.7±2.3) kg, P=0.000], visceral fat [(3.0±0.7) kg vs. (2.3±0.6)kg, P=0.000], visceral fat area [(97.6±22.2) cm~2 vs. (75.5±21.1) cm~2,P=0. 000], serum triglyceride [(1.98±0.94) mmol/L vs. (1.22±0.61) mmol/L, P=0.000]were all increased, while serum HDL [(1.23±0.32) mmol/L vs. (1.40±0.37) mmol/L, P=0.002]was decreased in NFLD group. The incidences of over-body fat (68.7% vs. 36.5%, P=0. 000),dyslipidemia (47.0% vs. 21.2%, P=0. 000), abdominal obesity (69.9% vs. 43.5%, P=0.001) and MS (49.4% vs. 9.6%, P=0.000) were obviously increased. But there were no statistical differences in serum TC [(4.93±0.94) mmol/L vs. (4. 73±1.07) mmol/L, P=0.219]and LDL [(3.23±0.80) mmol/L vs. (3. 07±0.89) mmol/L, P=0. 229]between the two groups. Logistic regression showed that high BMI (β=1.268, P=0.000, OR=3.56), over-total body fat percentage (β=0.902, P=0.023, OR=2.47)and the existence of MS (β=1. 664, P=0. 000, OR=5.28) were related to elderly type 2 diabetic patients complicated NFLD. Conclusions The high BMI, over-total body fat percentage are related to elderly type 2 diabetic patients complicated NFLD, and NFLD is probably one of components of metabolic syndrome.

ObjectiveTo study the efficacy and safety of heated humidified high flow nasal cannula ( HHHFNC) and nasal continuous positive airway pressure( NCPAP) ventilation for prevention of extubation failure in extremely low birth weight(ELBW)infants in our NICU.MethodsFrom Jan.2011 to Dec. 2014, 129 ELBW infants admitted to our hospital were randomly assigned into HHHFNC group and NCPAP group. The inclusion criteria were gestational age ( GA ) <34 w, birth weight ( BW )<1000 g, admission within 7 d after birth and transition to noninvasive respiratory support after a period of mechanical ventilation with an endotracheal tube. The primary outcome included:the incidence of extubation failure, nasal injury, air leak, abdominal distention and bronchopulmonary dysplasia ( BPD). Results Statistically significant difference sexisted between the two groups on oxygen the rapyduration, the time required reaching total enteral feedings and the incidences of nasal injury, air leak, abdominal & nbsp;distention and necrotizing enterocolitis ( P<0. 05). The incidence of extubation failure within 7 days was 25. 8℅ in HHHFNC group and 47. 6℅ in NCPAP group ( P <0. 05 ) . No differences between the 2 groups on total ventilation duration, non-invasive ventilation duration, re-intubation rate at 3d after extubation, BPD, retinopathy of prematurity ( ROP ) , intracerebral hemorrhage ( ICH ) , periventricular leukomalacia(PVL)and patent ductus arteriosus(PDA).Conclusions HHHFNC is an effective and safe method for prevention of extubation failure in ELBW infants.

Objective To compare anti-inflammatory and hepatoprotective effects of Bupleurum marginatum Wall. ex DC. and Bupleurum chinense DC. Methods The anti-inflammatory effect of two kinds of radix bupleuri decoction were compared using the xylene-induced ear edema mice model and the egg white-induced foot swelling rats model. Acute liver injury in mice was induced through intraperitoneal injection of peanut oil with 0. 1%carbon tetrachloride. Alanine aminotransferase ( ALT ) , aspartate aminotransferase ( AST ) , and alkaline phosphatase ( ALP ) in serum, superoxide dismutase ( SOD ) , malondialdehyde ( MDA ) , and glutathione ( GSH ) in liver tissue homogenate were detected. Histopathological changes, transforming growth factor-β ( TGF-β) and nuclear factor-κB ( NF-κB) in hepatic tissues were also observed. Results Ear swelling experiment showed that the decoction of both Bupleurum marginatum Wall. ex DC. and Bupleurum chinense DC. had equal anti-inflammatory effects (P>0. 05). Similarly,the decoction of Bupleurum marginatum Wall. ex DC. and Bupleurum chinense DC. improved levels of ALT,AST,SOD,GSH,and MDA to the same extent compared with the model control group ( P<0. 05 or P<0. 01 ). There was no significant difference among all groups in ALP levels ( P > 0. 05 ). Histopathological and immunohistochemical experiments showed that expressions of TGF-β and NF- κB were similar between the two kinds of radix bupleuri. Conclusion Bupleurum marginatum Wall. ex DC. and Bupleurum chinense DC. have similar anti-inflammatory and hepatoprotective effects.

Objective To establish an ideal focal cerebral ischemia reperfusion model in monkeys.Methods Adult healthy rhesus monkeys(Macaca mulatta) 12 cases(male 6 and female 6),aged 4-7 years and weighted 4.8-7.5kg.were used in this study.The middle cerebral artery occlusion/reperfusion(MCAO/R) model was established by inserting a standard micro-balloon catheter intraluminally from the carotid common artery or femoral artery into the proximal segment of the middle cerebral artery(MCA).The regional cerebral blood flow of MCA was occluded by expanding the micro-balloon to cause ischemia,and withdrawing the micro-ballon catheter to reperfuse the MCA.The MCAO/R model was evaluated by angiography,magnetic resonance angiography(MRA),magnetic resonance imaging(MRI),tetrazolium chloride(TTC) staining and neurological behavoral function scores.Results By inserting a micro-balloon catheter intraluminally from the carotid common artery or femoral artery into the MCA,the micro-balloon catheter could be inserted into the MCA to occlude blood flow,and no image of MCA shown on TV screen.In MCA blood flow supplied area,magnetic resonance T1,T2 and DWI showed high signals,TTC staining showed cerebral ischemic infarction,and correspondly the monkeys showed neurological function disorders.This method used a simple operatire procedure had a high successful rate,and could be repeated.Conclusion We showed ideal method to establish the MCAO/R model in monkeys by inserting intraluminally a micro-balloon catheter into the MCA.

OBJECTIVETo compare the clinical effects between interspinous dynamic internal fixation with Coflex system and posterior lumbar interbody fusion in treating lumbar degenerative disease.

METHODSFrom Jan. 2007 to Jan. 2010, 42 patients with lumbar degenerative disease were treated with interspinous dynamic internal fixation with Coflex system (non-fusion group, 21 cases) and posterior lumbar interbody fusion (fusion group, 21 cases), including 19 males and 23 females with an average age of (46.5 +/- 8.5) years (range, 33 to 62 years). The range of course of disease was from 6 to 84 months with an average of 36 months. All pathological changes were single segment in L4.5 by X-ray films or CT scanning, among them, single lumbar disc herniation was in 14 cases,combined with lumbar spinal stenosis in 28 cases. The clinical effects of all patients were evaluated according to Japanese orthopaedic association (JOA, 29 points) and Oswestry disability index (ODI); range of motion (ROM) of lumbar segments were measured by X-ray films; operative time, intraoperative blood loss,hospital stay were compared between two groups.

RESULTSAll patients were followed up for 10 months at least. In non-fusion group, JOA increased from 13.8 +/- 3.7 preoperatively to 24.6 +/- 4.6 at final follow-up (P < 0.01); ODI decreased from 65.8 +/- 7.5 to 25.6 +/- 5.1 (P < 0.01), ROM improved from 4.2 +/- 0.7 to 5.3 +/- 0.6 at L4,5 (P < 0.01 ). In fusion group,JOA increased from 13.2 +/- 3.2 preoperatively to 24.5 +/-4.3 at final follow-up (P < 0.01); ODI decreased from 65.5 +/- 8.2 to 26.5 +/- 6.1 (P < 0.01); all ROM at L.4,5 had lose (decreased from 4.2 +/- 3.2 to 0). There were significant differences in the operative time, intraoperative blood loss, hospital stay between two groups (P < 0.01), in non-fusion group was respectively (71.2 +/- 12.8) min, (56.6 +/- 25.5) ml, (4.4 +/- 1.3) d and in fusion group was respectively (121.0 +/- 23.2) min, (135.8 +/- 19.8) ml, (12.6 +/- 2.4) d. No complications were found with the Coflex system.

CONCLUSIONInterspinous dynamic internal fixation with Coflex system in treating lumbar degenerative disease is better than that of posterior lumbar interbody fusion in some aspects. Coflex system have high safety, less trauma, and can reserve lumbar mobility, which is effective in preventing adjacent intervertebral disc degeneration.

Adult ; Female ; Humans ; Internal Fixators ; Intervertebral Disc Displacement ; surgery ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Prostheses and Implants ; Spinal Stenosis ; surgery

OBJECTIVETo investigate the effects of serum containing Gumibao (Chinese character: see text) on the proliferation and differentiation of osteoblast induced by dexamethasone.

METHODSOsteoblasts were extracted from skulls in newly born (within 24 hours) SD rats, and digested with collagenase. The first passage of cells were used for experiments. Cells were cultured in the medium containing different concentrations of dexamethasone (0, 10(-8), 10(-7), 10(-6), 10(-5) ,10(-4) mol/L). Alkaline phosphatase staining were carried out after 1 week and numbers of mineralized nodes with alizarin red staining were observed after 3 weeks. Accordingly, following the treatment of 10(-5) mol/L dexamethasone for 1 week, cells were cultured in the medium with serum containing Gumibao (Chinese character: see text). One week after Cumibao (Chinese character: see text) treatment, cells were stained with Alkaline phosphatase and collagen I and PCNA were examined by Western-blot. However, the observation of numbers of mineralized nodes with alizarin red stain required one more week.

RESULTSHigh concentration of dexamethasone could inhibit the expression of PCNA, collagen I, alkaline phosphatase and reduce the number of mineralized nodes of osteoblast, while serum containing Gumibao (Chinese character: see text) could reverse the inhibition.

CONCLUSIONHigh concentration of dexamethasone could inhibit the proliferation and differentiation of osteoblastic cells, while serum containing Gumibao (Chinese character: see text) could reverse the inhibition.

Animals ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen Type I ; analysis ; Dexamethasone ; pharmacology ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; Female ; Osteoblasts ; cytology ; drug effects ; physiology ; Proliferating Cell Nuclear Antigen ; analysis ; Rats ; Rats, Sprague-Dawley

Objective To construct and apply a cell line screening Mycobacterium tuberculosis (Mtb)-specific tetramers of CD4+α/β T cell receptor(TCR). Methods The β chains of HLA class Ⅱ (DR) were amplified from tuberculosis patients by PCR. The pMT-HLA-DRB expression vectors that carries the HLA-DR 13 chain and pMT-HLA-DRA-P expression vectors which carries the genes of HLA-DR α chain loaded with Mtb antigen were transfected into S2 cells with the method of calcium phosphate transfection. The expressed Mtb peptide/HLA-DR complexes were primarily identified by the method of cell immunohistochemistry. The cell lines expressing Mtb peptide/HLA-DR complexes were used to screen tetramers of CD4+ TCR by flow cytometry. Results S2 cell lines expressing Mtb peptide/HLA-DR complexes on the cell surface were obtained, two kinds of Mtb specific tetramers of CD4+α/β TCR were screened. Conclusion S2 cell lines expressing Mtb peptide/HLA-DR complexes on the cell surface provide the solid basis of the further research on the TCR tetramers and are helpful for exploring new diagnostic study methods about tuberculosis and developing new vaccines.

Objective:To assess the feasibility and effectiveness of transnasal ultra-thin gastrointestinal endoscope-guided ileus tube insertion for the treatment of adhesive intestinal obstruction in the elderly.Methods:Randomized controlled trial, a total of 81 elderly patients with adhesive intestinal obstruction were enrolled, with 46 receiving transnasal ultra-thin gastroscope-assisted tube placement(observation group)and 35 receiving conventional transoral gastroscope-assisted tube placement(control group). Tube insertion was carried out.Data on the time needed for tube placement, incidents of oral or nasal bleeding, rates of successful tube placement, physical signs of symptom relief and imaging signs of symptom relief were recorded and compared between the two groups.Results:The symptom remission rate within 3 d was 93.5%(43/46)in the observation group and 88.6%(31/35)in the control group, and the rate for imaging signs of symptom remission was 82.6%(38/46)in the observation group and 74.3%(26/35)in the control group.The differences in symptom remission and imaging signs of symptom remission within 3 d were not statistically significant between the two groups( χ2=0.144, 0.830, all P>0.05). In the observation group, the time needed for tube placement was(15.4±4.2)min, which was significantly shorter than that in the control group(21.3±3.1)min( t=6.984, P<0.01). The rate of successful tube placement was 100% in both groups.In terms of adverse reactions, the observation group had 1 patient with nasal bleeding, 2 with nausea and vomiting, and 1 with a sore throat, with an overall adverse reaction rate of 8.7%(4/46), while the control group had no patient with bleeding, 18 with nausea and vomiting, and 4 with a sore throat, with an overall adverse reaction rate of 62.9%(22/35). There was no statistically significant difference in nasal bleeding(Fisher's exact probability P=0.568)or a sore throat( χ2=1.559, P=0.212), but the differences in the incidence of nausea and vomiting as well as the incidence of total adverse reactions were statistically significant( χ2=23.694, 26.752, both P<0.01)between the two groups. Conclusions:Transnasal ultra-thin gastroscope-assisted tube placement can reduce discomfort such as nausea and vomiting during tube placement, the time needed for the procedure is short with a high success rate, and therefore has very good clinical value, especially for elderly patients.