Animals ; Dibutyl Phthalate ; toxicity ; Female ; Hippocampus ; cytology ; growth & development ; Male ; Mitochondrial Proton-Translocating ATPases ; metabolism ; Neurons ; drug effects ; metabolism ; Pregnancy ; Rats ; Rats, Wistar

AIM: To investigate the role of adrenomedullin (AM) in diabetic nephropathy. METHODS: We observed the changes in the expression and secretion of AM, TGF-? 1 in the cultured human mesangial cells under high glucose condition and the contents of the laminin and type IV collagen in the supernatants. The effect of intervention with AM was also observed. RESULTS: High glucose condition resulted in increase in the expression and secretion of AM TGF-? 1 laminin and type IV collagen. AM reversed the influence of high glucose on the cultured human mesangial cells. CONCLUSION: These results showed that high glucose condition is one of stimulating factors of AM and the renal protective action of AM may be associated with suppression of TGF-? 1 and reducing excessive accumulation of laminin and type IV collagen.

Objective To assess the effect of chloramine disinfection on the formation of drinking water disinfection by-products(DBPs )and their mutagenicity.Methods The bacterial indices,chloroform and carbon tetrachloride content in the finished water samples using liquid chlorine and chloramine disinfection respectively were determined in August,2001(plentiful water season)and in March,2002(low water season).The mutagenicities of organic extracts from the water samples were tested using Ames test.The levels of DBPs and the mutagenic activities of water samples treated with different disinfection methods were compared.Results The total count of bateria and coliform bacteria were0/ml and0/L respectively at free chlorine con-centration of about 1mg /L in the chloraminated drinking water,which decreased81%-84%compared with those of the chlori-nated drinking water samples.The carbon tetrachloride concetration were all

Objective:To Observe the effect of recombinant tissue-type plasminogen activator (rt-PA) in treatment of acute myocardial infarct.Methods:Two groups of patients were observed.The subjects in the control group (35 patients) were treated with routine intervention,while those in thrombolysis group (15 patients) were treated with rt-PA in addition to the routine intervention.Results:There is a better outcome in the thrombolysis group as compared with that in the control group,in terms of length of angina relief,degree and length of EKG changes as well as the time for serum enzyme changes.The mortality rate in the thrombolysis group was significantly lower than that in the control group (P＜0.01).Conclusion:The rt-PA for thrombolysis in acute myocardial infarct patients can reduce the infarct size and improve the cardiac function rapidly as well as increase the survival rate.

Objective To evaluate the role of autophagy in hippocampal neurons in cognitive dysfunction caused by sevoflurane anesthesia in juvenile rats.Methods One hundred four male SpragueDawley rats, aged 7 days, weighing 10-16 g, were randomly assigned into 3 groups using a random number table: control group (group C, n =8), sevoflurane anesthesia group (group S, n =48), and sevoflurane anesthesia + rapamycin group (group SR, n =48).Group C inhaled 60％ oxygen for 6 h.S and SR groups inhaled 3.6％ sevoflurane anesthesia for 6 h, and in addition, rapamycin 2 mg/kg was injected intraperitoneally at 1 h before of sevoflurane inhalation in group SR.The equal volume of phosphate buffer solution was given in C and S groups.At 1 h before sevoflurane anesthesia (T0) , immediately after the end of anesthesia (T1) , and at 12, 24 and 48 h after the end of anesthesia (T2-4) , 8 rats were randomly selected from S and SR groups to determine the expression of autophagy-related proteins microtubule-associated protein 1 light chain 3 (LC3) Ⅰ , LC3 Ⅱ and Beclin-1 in hippocampal neurons (by Western blot).The ratio of LC3 Ⅱ/LC3 Ⅰ was calculated.Morris water maze test was performed at 5 weeks after the end of anesthesia to assess the cognitive function.The escape latency, frequency of crossing the original platform, and duration of swimming spent at the target quadrant were recorded.Results Compared with the values at T0, the expression of LC3 Ⅱ and Beclin-1 was significantly down-regulated at T1-3 , and the ratio of LC3 Ⅱ/LC3 [was decreased in group S, and the expression of LC3 Ⅱ and Beclin-1 was significantly up-regulated at T1-4, and the ratio of LC3 Ⅱ/LC3 Ⅰ was increased in group SR (P＜0.05).Compared with group C, the escape latency was significantly prolonged, the frequency of crossing the original platform was reduced, and the duration of swimming spent at the target quadrant was shortened at 3-5 days in group S (P＜0.05) , and no significant change was found in the parameters mentioned above in group SR (P＞ 0.05).Compared with group S, the escape latency was significantly shortened, the frequency of crossing the original platform was increased, and the duration of swimming spent at the target quadrant was prolonged at 3-5 days, the expression of LC3 Ⅱ and Beclin-1 was up-regulated at T1-4 , and the ratio of LC3 Ⅱ/LC3Ⅰ was increased in group SR (P＜0.05).Conclusion Weakened autophagy in hippocampal neurons is involved in cognitive dysfunction caused by sevoflurane anesthesia in the juvenile rats.

Objective To explore the shielding effect that Tween has made on the erythrocyte’ s surface antigen( ABO and D antigen mainly) and the stability of the RBC. Methods Various types red blood cells’ surface antigens were incubated with different concentrations of Tween,then the titers of RBC’ s surface antigen before and after incubation were compared. The erythrocyte’ s function alterations and sta-bility through the morphological obersavation,the osmotic fragility,RBC’ s own hemolysis rate,oxygen saturation( SO2 ) ,routine test of blood as well as the supernatant of free hemoglobin determination were confirmed. Results The masking effect of the Tween-20 on D antigen with the weaken titer keeping above“ +” was better and more stable than that on A,B antigen. The impact on B antigen was a little worse,and A was the worst. The advantaged concentration is about 0. 004%. Besides,the influence of Tween-80’s on B and D antigen was not apparent e-nough. Among all of the concentration,0. 74% and 0. 80% did a relatively better job,which also could keep the weaken titer above “ +”. Conclulsion The shielding effect that Tween has made on the erythrocyte’ s surface antigen( ABO and D antigen mainly) is stable,which keeps above “ +”. The morphology and function of the RBC does not change.

Objective: To explore the mechanism of An-pressing manipulation in improving post-stroke muscle spasticity, by observing the changes of γ-aminobutyric acid (GABA) and glycine (Gly) in plasma and gray matter of L1-L3 spinal cord anterior horn in post-stroke rats with muscle spasticity after An-pressing manipulation intervention. Methods: Ten of 80 adult male Sprague-Dawley (SD) rats were randomly selected as the blank group, and the remaining 70 were used for modeling. The middle cerebral artery occlusion (MCAO) rat model was established by insertion suture occlusion method in the left external carotid artery. Thirty rats with a Longa neurological score of 2-3 points and a modified Ashworth spasticity scale score of 1-, 1+, or 2 were included in the experiment. Using the random number table method, the 30 successfully modeled rats were randomly divided into a model group, an An-pressing tendon group and an An-pressing muscle belly group. Two days after modeling, rats in the An-pressing tendon group and An-pressing muscle belly group received An-pressing manipulation on the tendon and belly of quadriceps femoris muscle respectively, with the pressure of (350±50) g and the frequency of 5 s/time, 15 min per session, once a day for 5 continuous days. After the 5th treatment, the tension of the rat quadriceps femoris muscle was evaluated using the modified Ashworth spasticity scale. The Gly levels in rat plasma and L1-L3 segments of spinal cord were determined by enzyme-linked immunosorbent assay (ELISA). The GABA levels in rat plasma and L1-L3 segments of spinal cord were measured by high performance liquid chromatography (HPLC). Results: The decrease in rat muscle tension scored by the modified Ashworth spasticity scale in the An-pressing tendon group was more significant than that in the An-pressing muscle belly group (P<0.01); the increases in Gly and GABA levels in the rat plasma and L1-L3 segments of spinal cord were more significant in the An-pressing tendon group than those in the An-pressing muscle belly group (all P<0.01). Conclusion: Based on the theory of 'anti-stretch reflex' of tendon organs, the use of An-pressing manipulation to induce the 'anti-stretch reflex' by stimulating the tendon organs can improve the muscle spasticity of rats, which is better than An-pressing the muscle belly. Increased levels of Gly and GABA in rat plasma and L1-L3 segments of spinalcord may be one mechanism of An-pressing manipulation to improve muscle spasticity by stimulating tendon organs.

Objective To explore the anatomical basis,safty and manipuility of the puncture technique through anterolateral cervical approach.Methods Twenty two embalmed cadavers and 50 patients who underwent operation through anterolateral cervical approach were used to observe the cervical anatomical characteristics.During the cadaver dissection and operation,the following information was observed:the movement of the carotid sheath while the visceral sheath was pushed left,the features of interspace between the two sheaths at each cervical level,the movement of the esophagus and the interfascial space in which the esophagus moved while the trachea was pushed medially,and the movement of the carotid sheath while it was pushed medially and laterally.From January 2007 to December 2011,206 patients diagnosed as cervical discogenic pain or cervical vertigo were treated by radiofrequency using puncture technique through anterolateral cervical approach,including 93 males and 113 females,aged from 22 to 71 years (average,48years).A total of 434 discs were involved.Results Both in cadaver dissection and in operation,the following results were observed:there was no natural interspace between the carotid sheath and visceral sheath below the C4 level,as a result,manual separation was needed to create a interspace between the two sheaths for the puncture; the two layers of the prevertebral fascia could be separated easily; keeping the visceral sheath intact,the esophagus moved following the trachea when the latter was pushed medially,and the movement of the visceral sheath relative to the cervical vertebra occurred between the two layers of prevertebral fascia; the carotid sheath was primarily constituted by continuation of the prevertebral fascia,as a result,the movement of the carotid sheath was limited by its own tension and the prevertebral fascia.A total of 434 cases of puncture to cervical disc were performed smoothly with no complications related to the carotid artery,esophagus and cervical disc.Conclusion The puncture technique through anterolateral cervical approach requires a correct,thorough understanding of the anterior cervical anatomy,based on which this technique is safe and feasible.

Objective To study the effects of mild hypothermia on cardiomyocyte contractility improvement after ischemia-repeffusion injury and on the preservation of well-functioning mitochondrial respiratory capability.Methods A total of 50 newborn SD rats 1 ～ 2 days after delivery were sacrificed and their hearts taken to preserved in 4 ℃ cold D-hanks buffer solution with 0.12％ pancreatic proteinase and collagenase and then processed with 37 ℃ water bath to collect the cardiomyocytes cultured in DMEM medium with 10％ FBS for 5 days.The cardiomyocytes of rats were subjected to ischemia/reperfusion,in vitro,by oxygen and glucose deprivation(OGD)/oxygen and glucose restoration(OGR).The cardiomyocytes of rats after ischemia/reperfusion were divided into three groups:control group,hypothemia group and normothermia group.Contractile frequency and velosity were determined before OGD and 0 h,0.5h,1 h,1.5 h and 2 h after OGR.Ultrastructure changes of cardiomyocytes and mitochondrion were observed under transmission electron microscope(TEM)0 h and 2 h after OGR as well as assessment ot respiratory rate and respiratory control rate(RCR)with Clark oxygen electrode in each group.All data were analyzed with statistical software of SPSS 13.0.Results Contractile function of cardiomyocytes in hypothermia group and normothermia group declined to nadir at 0 h after OGR(P =0.000)and the contractile function of cardiomyocytes in hypothermia group was improved one hour later,compared with the normothermia group(P =0.000).Obvious swelling of mitochondrion was observed under TEM in normothermia group with little alteration after OGR.The RCR assessments indicated respiratory function in normothermia group was impaired after OGR(P =0.000)and this may be responsible for contractility dysfunction.Conclusions Mild hypothemia used after ischaemia can optimize the contractility of cardiomyocytes after a normothermia OGR,and the well-functioning respiratory capability of mitochondrion may be preserved in this process.

Objective To study the radiosensitivity enhancement effects of recombinant adenovirus mediated retinoblastoma 94 gene on the growth of esophageal carcinoma cells EC109.Methods EC109cells was transfected with recombinant Rb94 gene adenovirus and irradiated by 137Cs γ-rays.The cohorts were divided into groups as blank control,Ad-LacZ,Ad-Rb94,radiation and Ad-Rb94 combined with radiation.Cell inhibition ratio,cell cycle and expression of retinoblastoma protein of EC109 cells were analyzed.Results The growth of EC109 cells transfected with Ad-Rb94,radiation and Ad-Rb94 combined with radiation group was all inhibited.The group of Ad-Rb94 combined with radiation resulted in greater inhibition of cells growth compared with Ad-Rb94 group and radiation group ( F =23.31,P ＜0.05 ).Cells of G2 phases of EC109 cancinoma cells for Ad-Rb94 combined with radiation group were the highest,which was 50％.The combination of Ad-Rb94 and radiation group resulted in the greatest expression of retinoblastoma protein,which reached 71％,significantly higher than Ad-Rb94 infection and radiation groups ( x2 =8.31,6.73,P ＜ 0.05 ).Conclusions Retinoblastoma 94 gene combined with ionizing radiation can enhance the radiation sensitivity of EC109 cells.