Objective To observe the neural protective subsidiary effect of Wenyang Yiqi method on patients with severe traumatic brain injury under mild hypothermia therapy.Methods A prospective study was conducted in which 53 patients with severe traumatic brain injury treated by mild hypothermia were randomly divided into control group (26 cases) and observation group (27 cases). All the patients in the two groups received conventional western treatment combined with mild hypothermia therapy. In the observation group, additionally was given the representative drug of Wenyang Yiqi method, Shenfu injection 100 mL mixed into 5% glucose 500 mL intravenous drip once a day. At the end of mild hypothermia, the Shenfu injection was stopped. After treatment, the changes of intracranial pressure (ICP) on 1 (the day the treatment began), 2, 3, 4, 5, 6 and 7 days and the indexes levels, including S-100B, lactate dehydrogenase (LDH) and creatinkinase (CK) in the cranial spinal fluid (CSF) before treatment and 2, 4, 6 days after treament (CSF) were observed. The Glasgow coma score (GCS) before treament and 3, 5, 7, 14, 28 days after treament, and Glasgow outcome scale (GOS) on 28 days and 3 months after treatment were recorded, and the incidences of complications were calculated at the end of therapy.Results After treatment with the prolongation of therapeutic time, the levels of ICP were gradually increased in two groups and reached the peak values on the 4th day, then beganto fall, and on the 5th day it was significantly lower in observation group than that in control group [ICP (mmHg, 1 mmHg = 0.133 kPa): 16.11±1.23 vs. 18.73±1.42], persisting the same situation to the 7th day (14.17±0.80 vs. 16.94±1.00,P < 0.05). The levels of S-100 B in the two groups were progressively decreased after the treatment, on the 2nd day it was significantly lower in observation group than that in control group (μg/L: 1.21±0.43 vs. 1.86±0.57, P < 0.05), also persisting to the 6th day (0.40±0.09 vs. 0.94±0.15,P < 0.05); the levels of LDH and CK reached the peak values on the 2nd day, then began to fall, they were significantly lower in the observation group than those in the control group on the 4th day [LDH (U/L): 63.43±12.21 vs. 80.11±14.34, CK (U/L): 52.41±14.14 vs. 88.37±12.21, bothP < 0.05], and on the 6th day still there were statistically significant differences between the two groups. The GCS before treatment showed no statistically significant difference between the two groups (P > 0.05); after treatment, the GCS score of the two groups was progressively improved, and on the 14th day the score in the observation group began significantly higher than that in the control group (11.74±1.24 vs. 9.41±2.11,P < 0.05), persisting the same situation to the 28th day (12.68±2.51 vs. 10.67±1.99,P < 0.05). On the 28th day after treatment, the GOS showed no statistically significant difference between the two groups (2.35±0.16 vs. 2.43±0.22,P > 0.05), but the score in the observation group was significantly higher than that in the control group after treatment for 3 months (4.11±0.38 vs. 3.72±0.41, P < 0.05). The incidences of complications in the observation group were significantly lower than those in the control group [respiratory failure: 25.9% (7/27) vs. 50.0% (13/26), shock: 18.5% (5/27) vs. 53.8% (14/26), acute pulmonary edema: 14.8% (4/27) vs. 30.8% (8/26), stress ulcer: 22.2% (6/27) vs. 57.7% (15/26), hypoproteinemia: 40.7% (11/27) vs. 73.1% (19/26), allP < 0.05].Conclusion Wenyang Yiqi method has the subsidiary neural protective effect on patients with severe traumatic brain injury treated by mild hypothermia, and can improve their outcome.

BACKGROUND: Isoflavone isolated from Trifolium pratense L. has been found to be able to effectively inhibit bone resorption, reduce bone turnover rate, improve osteocyte activity and bone mineral density by enhancing the effect of estrogen, which is helpful for the prevention and treatment of osteoporosis. OBJECTIVE: To investigate the effect of Trifolium pratense L. extracts on the bone resorption and differentiation of osteoclasts.METHODS: Rat bone marrow cells were extracted, isolated by lymphocyte separation and cultured for 5 hours; then, the non-adherent cells were selected followed by induced by 30 μg/L macrophage colony stimulating factor and 75 μg/L RANKL (control groups), or different concentrations of Trifolium pratense L. extracts (0.3, 0.6 and 1.2 g/L) to observe their effect on the osteoclast differentiation and bone resorption. The levels of osteoclast differentiation-associated proteins c-fos and NFATcl were determined by western blot assay. RESULTS AND CONCLUSION: Compared with the control group, different concentrations of Trifolium pratense L. extracts could suppress osteoclast differentiation and bone resorption to different degrees. Tartrate-resistant acid phosphatase staining showed that Trifolium pratense L. extracts could significantly reduce the number of osteoclasts. Western blot assay results suggest that Trifolium pratense L. extracts significantly inhibited the expression levels of c-fos and NFATcl. These results reveal that Trifolium pratense L. extracts can inhibit osteoclast differentiation and bone resorption.

BACKGROUND:Ischemia/reperfusion (IR)injury during the pancreas transplantation can cause numerous postoperative complications, among which,secondary pancreatitis can cause small intestinal mucosal injury and result in severe Consepuence.OBJECTIVE:To observe the protective effect of ischemic preconditioning (IPC) on small intestinal mucosal barrier after pancreas transplantation in rats.DESIGN:Randomized controlled animal trial.SETTING:Department of Gastrointestinal Surgery,Xijing Hospital,Fourth Military Medical University of Chinese PLA.MATERIALS:This trial was done in the Laboratory of Gastrointestinal Surgery,Xijing Hospital,Fourth Military Medical University of Chinese PLA between September 2001 and April 2004.Eighty-three male SD rats were involved in this trial.METHODS: Forty-seven rats were randomly chosen to prepare diabetic rat models by penile-intravenous injection of 65 mg/kg streptozotocin.Thirty-six successful model rats were randomized into 3 groups,with 12 in each group:IR group,donor IPC(DIPC)group and recipient with two hindlims IPC(RIPC)group.Twelve of the remaining 36 normal rats served as control group,and the other 24 rats were used as donors.Laparotomy was conducted only in control group,and pancreas transplantation was conducted in the other 3 groups In DIPC group,the splenic vessels of donors were blocked for 5 minutes and reperfused for 5 minutes twice before obtaining pancreas from donor;In the RIPC group, blood flow of two hindlimbs of recipients was blocked for 5 minutes and reperfused for 5 minutes before reperfusing the pancreas of donor,and this procedure was repeated 3 times.IR group was untouched.MAIN OUTCOME MEASURES:① On the 5th day after operation,6 rats were randomly chosen from each group to detect small intestinal permeability[expressed with plasm fluorescent-isothiocyanate-dextran(FITC-dextran)concentration]and absorption function(expressed with plasm xylose concentration).② On the 5th day after operation.blood was taken from the left 6 rats in each group to detect serum tumor necrosis factor-α(TNF-α) and nitric oxide(NO)level as well as superoxide dismutase(SOD)and amylase activity.Ileal mucosal tissue was taken to detect wet weight of small intestinal mucosa,the height and width of microvilli,malonaldehyde(MDA)level and myeloperoxidase(MPO)activity.At the same time,mesenteric lymph node,liver and splenic tissue were taken to perform bacterial culture.Bacterial translocation was observed.RESULTS:After supplement,72 rats were involved in the result analysis.①Plasm FITC-dextran concentration of IR group were higher than that in control group,DIPC group and RIPC group,respectively(P＜0.01).②Plasm xylose concentration in the IR group was lower than that in the control group,DIPC group and RIPC group,respectively(P＜0.01).③Bacterial translocation rate in the IR group was higher than that in the control group,DIPC group and RIPC group,respectively(P＜0.01).④Small intestinal mucosal injury degree in the IR group was lower than that in the other 3 groups(P＜0.01).⑤Small intestinal MPO activity and MDA level in IR group were significantly higher than those in the other 3 groups(P＜0.01). Serum SOD activity and NO level were lower but amylase activity and TNF-α 1evel were higher in the IR group as compared with the other 3 groups(P＜0.01).CONCLUSION:IPC of two hindlimbs in both donor and recipient can protect small intestinal mucosal barrier and reduce bacterial translocation rate after pancreas transplantation in rats.

Objective To study the effect of chronic fluorosis on neurobehavioral development,the ability of learning and memory in offspring of rats,and the antagonistic effect of antioxidant Vitamin E (Vit E).Methods According to body weight,forty-five 1-month-old Sprague-Dawley(SD) rats of 30 females and 15 males were divided into three groups by random number table,including control group,fluorosis and Vit E antagonistic groups (15 rats with 10 females and 5 males in every group).Five months after establishing the animal model with chronic fluorosis and Vit E gavage treatments (fluoride ＜ 1,50,50 mg/L,respectively; Vit E 0,0,50 mg/kg,respectively),the rats were mated in 2:1 proportion of female:male in different groups,respectively.The fertility index of female and neurobehavioral development indicators in offspring were observed.Spatial learning and memory of offspring after birth for 30 d were evaluated by using Morris Water Maze test.Results The female fertility index exposed to fluorosis and Vit E were not significantly different as compared to those of control group(all P ＞ 0.05) ; in contrast to control groups[(6.4 + 1.8),(15.1 + 1.7)d],the time that completed the surface righting reflex [(8.1 + 1.4),(7.9 + 1.5)d] and the air righting reflex [(17.7 + 2.3),(17.2 + 1.8)d] were delayed in the offspring in fluorosis and Vit E antagonistic groups(all P ＜ 0.05) ; the completed avoidance precipice reflex and the auditory consternation did not changed significantly(all P ＞ 0.05); In addition,compared with control and Vit E antagonistic groups [(31.74 + 17.78),(34.97 ± 15.44)s,(4.50 ± 2.51),(3.80 ± 1.87)time],the average escape latency and exploration platform at five days were decreased in 30 d offspring of fluorosis group[(42.03 + 16.45)s,(2.20 + 1.62)time].Conclusion Neurobehavioral development as well as learning and memory ability in rat offspring are impaired by long-term exposure to fluoride and Vit E has exhibited an antagonistic effect to the toxicities of fluoride.

Objective To study the correlation between the features of peripheral lung carcinoma on spiral CT(SCT) and bFGF expression and angiogenesis. Methods 22 cases of peripheral lung carcinoma proved pathologically and examined with CT(SCT) both plain scan and contrast-enhanced scan were analyzed retrospectively . The expressions of bFGF and angiogenesis were detected with immunohistochemical SP method. The SCT features were compared with immunohistochemical results such as bFGF expression, microvessel density (MVD). Results The positive expression of bFGF was 59.9%, and the mean MVD was 46.10?18.18. There was significantly positive correlation between bFGF and MVD (P

Objective To observe the expression of NF-E2-related factor 2 (Nrf2) and Kelch-like ECH-associated protein-1 (Keap1) at mRNA and protein levels in the brain of rats with chronic fluorosis and to reveal the mechanism of brain damage induced by the factors.Methods Ninety SD rats were divided into three groups (30 rats in each group,half male half female) by the random number table method according to body weight.The control group was fed with normal tap-water,high-fluoride group with 50 mg/L fluoride (NaF) added in drinking water;and the high-fluoride plus vitamin E (Vit E) group with the same dose of NaF as the high-fluoride group,but giving 5 mg/kg Vit E by intragastric administration.The experiment period was 10 months.The fluoride contents in urine and bone were detected by fluoride-ion selective electrode.The protein and mRNA levels of Nrf2 and Keap1 in brain of rats were detected by Western blotting and quantitative real time PCR,respectively.The activity of superoxid dismutas (SOD) and the content of lipid peroxidation (MDA) were measured by biochemistry methods.Results Dental fluorosis was detected in high-fluoride group.The differences of fluoride contents in urine and bone were statistically significant between groups (F =6.87,182.87,all P ＜ 0.05).The urine fluoride [(2.16 ± 0.39),(2.07 ± 0.15)mg/L] and bone fluoride [(211.07 ± 40.52),(82.09 ± 28.60)mg/kg] in the high-fluoride and high-fluoride plus Vit E groups were higher than those of the control group [(1.70 ± 0.24)mg/L,(34.67 ± 11.15)mg/kg,all P ＜ 0.05].The differences of mRNA and protein levels of Nrf2 and Keap1 in brains of rats,SOD activity,MDA content were statistically significant between groups (F =654.33,432.87,447.45,398.88,68.34,68.34,all P ＜ 0.05).The mRNA levels of Nrf2 and Keap1 [(320.18 ± 6.83)％,(267.37 t 7.22)％] were increased compared to those of control group [(100.00 ±3.00)％,(100.00 ± 2.75)％,all P ＜ 0.05];the protein levels of Nrf2 and Keap1 [(283.28 ± 6.89)％,(196.32 ± 5.57)％]were also raised compared with those of control group [(100.00 ± 8.71)％,(100.00 ± 9.23)％,all P ＜ 0.05];the activity of SOD [(22.10 ± 2.10)μ,mol/kg] in brain of rats in fluoride group was significantly lower and the content of MDA [(8.63 ± 0.77) μmol/kg] was higher than those of control group [(35.05 ± 2.98),(1.25 ± 0.64) μmol/kg,all P ＜ 0.05].In high-fluoride plus Vit E group,the mRNA levels of Nrf2 and Keap1 [(243.23 ± 5.34)％,(180.54 ± 4.48)％] and the protein levels of Nrf2 and Keap1 [(210.88 ± 4.79)％,(150.68 ± 6.49)％] were lower than those of high-fluoride group (all P ＜ 0.05);the activity of SOD [(26.33 ± 1.84)μmol/kg] was significantly higher and the content of MDA [(4.88 ± 0.84)μmol/kg] was lower than that of high-fluoride group (all P ＜ 0.05).Correlation analysis showed that increased levels of Nrf2 and Keap1 were positively correlated with the level of MDA in rat brain (r =0.69,0.33,all P ＜ 0.05),but negatively correlated with the activity of SOD (r =-0.78,-0.80,all P ＜0.05).Conclusion The expression of Nrf2 and Keap1 in brain of rats with fluorosis is significantly increased and positively correlated with the content of fluoride in bone and the level of oxidative stress,whereas vit E can attenuate these abnormal changes induced by fluoride,which might be one of the mechanisms of brain damage of the disease.

Objective To detect the levels of oxidative stress in brain and serum of rats with chronic fluorosis and the antagonistic effects of vitamin E (VitE),and to reveal the role of oxidative stress in brain injury.Methods Thirty healthy SD rats were divided into three groups based on body weight by means of a random number table (10 rats in each group,half male and half female).In the control group,the rats were fed with drinking water containing less than 0.5 mg/L fluoride;in the fluoride group,the rats were fed with high doses of sodium fluoride in drinking water (50.0 mg/L) and the VitE antagonistic group were fed with the same content of fluoride in drinking water as the fluoride group,but adding VitE (50.0 mg/kg) by intragastric administration once a day.All rats were fed with normal diet (6.2 mg/kg).After exposure to fluoride for 10 months,all rats were put to death,dental fluorosis of the rats was examined and the fluoride content in bone was determined by fluoride-ion selective electrode;the activity of superoxide dismutase (SOD) was determined by the xanthine oxidase method and glutathione peroxidase (GSH-Px) by the colorimetric method,the level of malonaldehyde (MDA) by the glucosinolates barbituric acid fluorescence method and the levels of OH-,H2O2 and O-·2 in rat serum and/or brain were detected by the colorimetric method.Results In the rats of the fluoride group,fluoride content in bone was higher as compared to control [bone fluoride:(211.07 ± 48.52) vs.(33.40 ± 9.26) mg/kg,P ＜ 0.01].The activities of SOD and GSH-Px in rat brains of the fluoride group [(20.10 ± 1.98) kU/g,(28.70 ± 19.35) kU/L] were significantly lower than those of controls [(37.05 ± 3.13) kU/g,(59.63 ± 12.83) kU/L,all P ＜ 0.01],the activity of SOD in VitE antagonistic group [(26.27 ± 1.74) kU/g] was higher than the fluoride group (P ＜ 0.01);the activities of SOD and GSH-Px in rat serum of the fluoride group were significantly decreased [(11.55 ± 1.75) kU/L,(79.50 ± 19.18) U/L] than those of controls [(20.79 ± 2.43) kU/L,(170.00 ± 14.68) U/L,all P ＜ 0.01],the activity of SOD in VitE antagonistic group [(17.23 ± 0.68) kU/L] was higher than the fluoride group (P ＜ 0.01).The levels of MDA in rat brain and serum of the fluoride group [(8.84 ± 0.69) μmol/L,(1.46 ± 0.11) nmol/L] were significantly higher than those of controls [(1.27 ± 0.74) μmol/L,(0.83 ± 0.10) nmol/L,all P＜ 0.01],VitE antagonistic groups [(4.51 ± 1.13) μmol/L,(1.29 ± 0.02) nmol/L] were lower than the fluoride groups (all P ＜ 0.01).The levels of OH-,H2O2 and O-·2 in rat brains of the fluoride group [(24.24 ± 1.80) kU/g,(15.28 ± 2.97) mmol/L,(6.53 ± 0.96) U/g] were significantly higher than those of controls [(11.44 ± 1.63) kU/g,(5.28 ± 1.20) mmol/L、,(2.93 ± 0.42) U/g,all P ＜ 0.01],VitE antagonistic groups [(14.43 ± 0.76) kU/g,(8.09 ± 0.55) mmol/L,(4.41 ± 0.49) U/g] were lower than the fluoride groups (all P ＜ 0.01).Conclusions Elevated levels of oxidative stress are found in brain and serum of the rats with chronic fluorosis,which may be a main mechanism of brain injury.VitE may play an important antagonistic role in oxidative damage induced by fluoride toxicity.

Objective To investigate the correlation of the expression of vascular endothelial growth factor(VEGF) and microvessel density(MVD) value with spiral CT(SCT) features in lung carcinoma.Methods Pre-and post-enhanced SCT scans were performed in 41 cases of lung carcinoma proved surgically and pathologically.There were 29 men and 12 women.22 cases were peripheral lung carcinoma,and 19 cases were central lung carcinoma.SCT characteristics of all cases were analyzed and recorded.SP immunohistochemical technique was experimented on each pathologic specimen.The enhanced range of lesions and the expression of VEGF,MVD value were analyzed using stepwise regression.Chi-square test was performed to determine the correlation between the expression of VEGF,MVD value and CT features in peripheral lung carcinoma.Results CT enhanced degree of lesions increased with the increased level of positive expression of VEGF and MVD value by stepwise regression analysis(standardized coefficients were 0.498 and 0.399,respectively,P3cm in diameter were significantly higher than those of the group with tumor size ≤3 cm.The MVD value increased significantly in cases with deep lubulation sign,spiculate protuberance and vessel convergence(P

Objective To investigate the changes of expression of quinone oxidoreductase-1 (NQO1) and heme oxygenase-1 (HO1) at protein and mRNA levels in the brains of rats with chronic fluorosis,effect on NF-E2-related factor 2/antioxidant response element (Nrf2/ARE) signal pathway,and reveal the mechanism of brain damage induced by the disease.Methods SD rats were randomly divided to two groups of 30 each (half females and half males),e.g.the normal control group (drinking water containing less than 0.5 mg/L of fluorine) and fluoride exposed group (drinking water containing 50.0 mg/L sodium fluoride,NaF).All rats were examined at the 10 months after feeding NaF.Dental fluorosis of rats was observed; the fluoride contents in urine and bone were detected by fluoride-ion selective electrode; protein and mRNA levels of NQO1 and HO1 in brains were detected by Western blotting and quantitative real timePCR,respectively.Results The dental fluorosis was observed,and contents of fluoride in urine [(2.16 ± 0.39)mg/L] and bone [(211.07 ± 40.52)mg/kg] determined in the rats of the fluoride group were higher than those of controls [(1.70 ± 0.24)mg/L,(34.67 ± 11.15)mg/kg,t =2.11,3.23,all P＜ 0.05].The protein expression levels of NQO1 and HO1 in the brains of rats with fluorosis [(255.2 ± 14.3) ％ and (187.2 ± 11.1)％] were also higher than those of controls [(100.0 ± 12.2)％,(100.0 t 8.9)％,t =2.14,2.05,all P ＜ 0.05]; the mRNA levels of NQO1 and HO1 [(210.2 ± 9.8)％ and (154.5 ± 7.4) ％] in the rats of the fluoride group were increased as compared to those of controls [(100.0 ± 10.4)％,(100.0 ± 9.7)％,t =2.33,2.75,all P ＜ 0.05].Conclusion The expression of NQO1 and HO1 in brain of rats with fluorosis are significantly increased,which may be due to the activation of Nrf2/ARE signal pathway and may play a compensative role in enhancing antioxidant ability.

Objective To detect the expression of muscarinic acetylcholine receptors (mAChRs) at mRNA and protein levels in the brain of rats with chronic fluorosis and to reveal the role of the receptors in brain injury and learning and memory deficits.Methods Sixty healthy SD rats were divided into two groups (30 rats in each group,half males and half females) by random number table method according to body weight.In the control group,the rats were fed with drinking water containing no more than 0.5 mg/L fluoride; in the fluoride group,the rats were fed with high doses of sodium fluoride in drinking water (50.0 mg/L).Each group was fed with normal diet (6.2 mg/kg).After being exposed to fluoride for 10 months,behavioral performance was measured with Morris water maze,including the escape latency time and the numbers of crossing platforms.After being sacrificed,rat brains were taken and weighted.M1 and M3 subunits at mRNA and protein levels were detected by real-time PCR and Western blotting,respectively; the correlation between protein levels of the receptor subunits and the ability of learning and memory was analyzed.Results In fluoride group,the escape latency time [(21.68 ± 2.90)s] was significantly longer than that of control group [(6.14 ± 1.71)s,t =0.289,P ＜ 0.05]; and the number of crossing platforms [(11.62 ± 2.26)times] was significantly decreased as compared to that of control group [(19.00 ± 3.69)times,t =0.352,P ＜ 0.05].Furthermore,the mRNA expression [(17.07 ± 6.89)％,(12.25 ± 5.03)％] and the protein levels [(71.07 ± 6.89)％,(32.25 ± 4.66)％] of M1 and M3 receptors in rat brains were significantly lower as compared to those of controls [(100.00 ± 3.00)％,(100.00 ± 2.15)％ and (100.00 ± 9.01)％,(100.00 ± 10.33)％,t =0.210,0.157,0.095,0.296,all P ＜ 0.05].The escape latency and M1,M3 protein levels were negatively correlated (r =-0.683,-0.700,all P ＜0.05),and the number of space exploration and M1,M3 protein levels were positively correlated (r =0.867,0.837,all P ＜ 0.05).Conclusion Declined expression of mAChRs at mRNA and protein levels have been detected in the brain of rats with chronic fluorosis,which may be one of the main mechanism concerning the learning and memory deficits.