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.

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.

Objective To observe the changes of learning and memory ability and detect the expression of muscarinic acetylcholine receptor (mAChR,M receptor) at mRNA and protein levels in brains of offspring rats with chronic fluorosis,and to reveal the mechanism of the central nervous system damage.Methods Forty healthy SD rats were divided into two groups (20 in each group,half male and half female) by random number table 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 dose of sodium fluoride in drinking water (50.0 mg/L fluoride).Each group was fed with normal diet (6.2 mg/kg fluoride).After exposed to fluoride for 6 months,each group was mated,and brains of newborn offspring rats aged 1,7,14,21 and 28 days were taken,and expression of M1 and M3 receptors at mRNA and protein levels were analyzed by real-time PCR and Western blotting,respectively.Behavioral changes were measured by Morris water maze test at the 28 days after birth.The correlations between protein levels of M1 and M3 receptors and the ability of learning and memory at the 28 days after birth were analyzed.Results In fluoride group of the offspring rats at 28 days after birth,the escape latency time [(35.61 ± 9.00)s] was significantly longer than that in control group [(8.46 ± 3.09)s,P ＜ 0.05],while the numbers of crossing the platforms and the time of staying the platforms [(5.00 ± 2.90)times,(16.66 ± 2.79)s] were significantly decreased as compared to that of control group [(15.17 ± 3.66)times,(22.51 ± 2.66)s,all P ＜ 0.05].Furthermore,the mRNA expression and the protein levels of M1 and M3 receptors in rat brain at each phase in fluoride group were significantly decreased as compared to controls [M1 mRNA in control groups:(100.00 ± 11.00)％,(100.00 ± 17.57)％,(100.00 ± 9.14)％,(100.00 ± 7.52)％,(100.00 ± 15.78)％;M1 mRNA in fluoride groups:(20.47 ± 8.07)％,(14.00 ± 4.53)％,(16.57 ± 7.62)％,(25.56 ± 12.78)％,(16.27 ± 4.82)％;M3 mRNA in control groups:(100.00 ± 16.30)％,(100.00 ± 14.40)％,(100.00 ± 7.20)％,(100.00 ± 14.31)％,(100.00 ± 13.16)％;M3 mRNA in fluoride groups:(29.17 ± 8.00)％,(12.77 ± 2.22)％,(26.40 ± 7.20)％,(15.74 ± 3.55)％,(28.14 ± 7.53)％;M1 protein in control groups:(100.00 ± 2.24)％,(100.00 ± 8.30)％,(100.00 ± 4.61)％,(100.00 ± 13.78)％,(100.00 ± 11.72)％;M1 protein in fluoride groups:(20.47 ± 8.07)％,(14.00 ± 4.53)％,(16.57 ± 7.62)％,(25.56 ± 12.78)％,(16.27 ± 4.82)％;M3 protein in control groups:(100.00 ± 16.30)％,(100.00 ± 14.40)％,(100.00 ± 7.20)％,(100.00 ± 14.31)％,(100.00 ± 13.16)％;M3 protein in fluoride groups:(29.17 ± 8.00)％,(12.77 ± 2.22)％,(26.40 ± 7.20)％,(15.74 ± 3.55)％,(28.14 ± 7.53)％,P ＜ 0.05 or ＜ 0.01].The escape latency and M1,M3 receptors protein levels were negatively correlated (r =-0.827,-0.742,all P ＜ 0.05),and the number of space exploration and M1,M3 receptors protein levels were positively correlated (r =0.843,0.806,all P ＜ 0.05).Conclusion The expression of M receptor at protein and mRNA levels in offspring rat brains of different ages are significantly declined,which might be one of the mechanism of the decreased ability of learning and memory 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 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 investigate the pharmacokinetics of D-polymannuronicate after single and successive administrations for 7 days by means of intravenous injection and intragastric administration.Methods Bioanalysis for the determination of D-polymannuronicate plasma concentration was applied in rats,and parameters of pharmacokinetics were calculated by DAS2.1.1 software.Results The well linearity(r=0.9991) in 0.05～150mg?L~(-1) of plasma concentrations.The recovery rate was between 94.72%～103.21%,the derivations withinday and between-days were less than 15%.Zata were 0.89,1.06,0.93,0.85 h~(-1) and t_(1/2z) were 0.78,0.69,0.75,0.87h corresponding with the four methods of administration.The bioavailabilities were 3%～5%.Conclusion The bioavailability was low.The ends of the elimination of two means of intravenous injection and intragastric administration were same. There was no significant difference in pharmacokinetic parameters of single and successive administrations.

Objective:To explore the etiology and characteristics of chronic wet cough in children in Qingdao.Methods:Patients with chronic wet cough treated at respiratory clinic of the Women and Children′s Hospital Affiliated to Qingdao University from July 2018 to June 2019 were included in this study.After three-month follow-up, the etiological data was analyzed.Results:(1)A total of 213 children were included, ranging in age from 1 month to 14 years old, including 38 cases of 1 month~1 year old, 47 cases of 1~3 years old, 87 cases of 3~6 years old, and 41 cases of 6~14 years old.The median age was 4.7 years.The top four causes of chronic wet cough in children were upper airway cough syndrome(33.8%), protracted bacterial bronchitis(20.7%), asthma with upper airway cough syndrome(15.5%), and asthma with infection(10.8%). Other causes were postinfection cough, pertussis syndrome, bronchiectasis, gastroesophageal reflux, bronchial foreign body, abnormal airway development, cystic fibrosis and so on.(2)The first cause of chronic wet cough in different age groups: 1 month to 3 years old group was protracted bacterial bronchitis; 3 to 14 years old group was upper airway cough syndrome.(3)The causes of chronic wet cough showed seasonal differences.Upper airway cough syndrome and cough after infection had a more balanced incidence throughout the year; protracted bacterial bronchitis and pertussis syndrome were common in winter; asthma with upper airway cough syndrome and asthma with infection were common in spring and autumn.Conclusion:Upper airway cough syndrome, protracted bacterial bronchitis, asthma with upper airway cough syndrome, and asthma with infection are the 4 leading causes for children with chronic wet cough in Qingdao.The causes of chronic wet cough have age and seasonal differences.

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.