BACKGROUND: It has been demonstrated that insulin-like growth factor-1 (IGF-1) is a kind of neurotrophic factor and protects from cerebral ischemia/reperfusion injury, the expression of IGF-1 is associated with the attack of ischemic stroke. The effects of IGF-1 and its receptor (IGF-1R) on neurobehavioral function are to be further studied.OBJECTIVE: To observe the effects of IGF-1 and IGF-1R on neurobehavioral function in rat models of cerebral ischemia/reperfusion injury.DESIGN: A randomized controlled observation.SETTING: Institute of Cerebrovascular Diseases, Affiliated Hospital of Qingdao University Medical College.MATERIALS: The experiments were carried out in Shandong Key Laboratory for Prevention and Treatment of Brain diseases. Twenty-eight healthy adult Wistar rats of clean degree, weighing 220-260 g, were provided by the experimental animal center of Shandong University.METHODS: The rats were randomly divided into experimental group (n =24) and sham-operated group (n =4). The middle cerebral artery occlusion/reperfusion (MCAO/R) models were established by inserting a thread through left external-internal carotid arteries. The sham-operated rats were given the same treatments except inserting thread. ①Neurologic deficit test: The rats in the experimental group were assessed according to Bederson standard after 1-hour ischemia and 6, 12-hour, 1, 3, 7 and 14-day reperfusion respectively. The sham-operated rats were assessed at corresponding time points; Without neurologic deficit was marked as 0 point; flexion of anterior claws as 1 point; unable to act against the pushing from the contralateral side as 2 points; circling while walking as 3 points; shaking as 4 points;unconscious mind as 5 points. ② Sample collection and treatment: The samples in the experimental group were collected after 1-hour ischemia and 6, 12-hour, 1, 3, 7 and 14-day reperfusion, and those in the sham-operated group ere collected at 24 hours postoperatively. The rats were anesthetized, brain samples were got at about 5 mm posterior to optic chiasma after brains were removed completely, then serial coronal sections (5 μm) were prepared, and 1 from 10 sections was stuck to the cover glasses treated with poly-L-lysine. ③ Morphological observation of neurons: The neurons in brain were observed by toluidine blue staining. ④ Detection of IGF-1 and IGF-1R: The expressions of IGF-1 and IGF-1R in cortex and striatum were detected with immunohistochemical technique, 4 fields were randomly selected to count the positive cells under high-power microscope (×400).MAIN OUTCOME MEASURES: ① The neurologic deficit; ② Morphological changes of neurons in brain; ③ Expressions of IGF-1 and IGF-1R in cortex and striatum.RESULTS: All the 28 rats were involved in the analysis of results. ① The neurologic deficit: The scores of neurologic deficit were (1.50±058) and (1.50±0.78) in rats after 7 and 14-day reperfusion, which were lower than that in rats after 6-hour reperfusion [(3.00±0.00), P ＜ 0.05]. ② Morphological changes of neurons in brain: The neurons in ischemic area appeared as paryopyknosis and became irregular in shape, there were obvious gaps around the cells, also deeply stained as purplish blue, nucleolus disappeared, and there were many scattered cellular fragments. ③ Expressions of IGF-1 and IGF-1R in cortex and striatum: The numbers of IGF-1 positive cells in cortex were (8.75±2.06), (11.13±1.14),(19.75±3.18), (17.38±3.11 ) and (11.23±2.28) respectively in rats after 6, 12-hours and 1, 3, 7-day reperfusion, which all were higher than that in sham-operated rats [(3.88±1.46), P ＜ 0.05], the numbers of IGF-1 positive cells in striatum were(8.25±2.21), (11.34±2.21), (18.23±2.64), (18.56±2.34) and (11.31±2.14) respectively in rats after 6, 12 hours and 1, 3, 7days reperfusion , which were also higher than that in sham-operated rats [(4.12±2.24), P ＜ 0.05]. The numbers of IGF-1R positive cells in cortex were (7.63±1.50), (10.50±2.34), (15.55±3.12), (15.37±3.01), (8.86±2.75) respectively in rats after 6, 12-hours and 1,3,7-day reperfusion, which all were higher than that in sham-operated rats [(4.13±1.81), P ＜0.05]. Those in striatum were (8.33±2.31), (10.24±2.09), (14.72±2.17), (14.24±2.77), (8.38±2.05), which were also higher than that in sham-operated rats [(3.76±2.35), P ＜ 0.05].CONCLUSION: The neurological function is damaged after cerebral ischemia/reperfusion, but it has a trend of self-recovery. The expressions of IGF-1 and IGF-1R are mainly distributed in cortex and striatum. Higher expressions of IGF-1 and IGF-1R maintain during 12 hours to 7 days after reperfusion and have a peak value at 1-3 days, which suggests that early expression of IGF-1 and IGF-1R are certain related to the recovery of neurological function.

BACKGROUND: Brain injury can induce the increased expression of basic fibroblast growth factor (bFGF) in brain,whereas FGFR is a very important player in the cell proliferation and differentiation, angiogenesis, skeletogeny, etc.OBJECTIVE: To observe the effect of bFGF and its receptor on neuronal apoptosis following cerebral ischemia/reperfusion injury in rats.DESIGN: A randomized grouping design and animal experiment.SETTING: Institute of Cerebrovascular Disease, Affiliated Hospital of Qingdao University Medical College.MATERIALS: Twenty-eight healthy adult Wistar rats of clean degree, weighing 220-260 g, were provided by the experimental animal center of Shandong University. Rabbit-anti-rat bFGF and fibroblast growth factor receptor-1(FGFR-1) monoclonal antibodies were provided by Wuhan Boster Biological Technology, Co.,Ltd.METHODS: The experiment was carried out in Shandong Key Laboratory for Prevention and Treatment of Brain diseases.① The rats were randomly divided into experimental group (n =24) and sham-operated group (n =4). Models of middle cerebral artery occlusion/reperfusion (MCAO/R) were established by thread occlusion via left external-internal carotid arteries, and 4 rats in the experimental group were sampled at 1-hour ischemia/6, 12-hour, 1, 3, 7 and 14-day reperfusion respectively. The rats in the sham-operated group were given the same treatment without inserting thread.After anesthesia, the brain was removed completely by cutting head, then the brain tissue at about 5 mm posterior to optic chiasma was cut down, then serial coronal sections (5 μm) were prepared. ② The brain tissues were stained with ematoxylin-eosin (HE), and the forms of neurons were observed under microscope. ③ TdT-mediated dUTP-biotin nick end labeling (TUNEL) method: there were buffy granules in nucleus which was positively stained (apoptosis). Four fields were randomly selected from cortex and striatum to count positive cells under high-power microscope (×400). ④ The sections were stained with rabbit-anti-rat bFGF and FGFR-1 monoclonal antibodies, 4 fields were randomly selected from cortex and striatum to count positive cells under high-power microscope (×400).MAIN OUTCOME MEASURES: Apoptosis and the expressions of bFGF and FGFR-1.RESULTS: All the 28 rats were involved in the analysis of results. ① In the experimental group, the neurons in the ischemic sites were obviously decreased, some neurons appeared as paryopyknosis and became irregular, also deeply stained as purplish blue, nucleolus disappeared, and there were many scattered cellular fragments. ② In the sham-operated group, there were a few apoptotic neurons in the brain tissue, and the apoptotic neurons were obviously increased after ischemia, which mainly observed in cortexes and striatums of frontal and paritetal lobes. In the experimental group, apoptotic cells in cortexes began to increase gradually at 6 hours, and there were more cells at 12hours and 3 days, which reached the peak value at 1 day, and began to decrease at 3 day, but there were still more apoptotic cells at 14 days than in the sham-operated group. The number of apoptotic neurons and the changing trend in striatums were generally the same as those in cortexes (P ＞ 0.05). ③ In the sham-operated group, there were weak bFGF expression in the neurons of brain tissue, but there were fewer lightly stained positive cells. After cerebral ischemia, the bFGF expressions were increased, mainly observed in cortexes and striatums. The bFGF expression appeared at 6 hours after cerebral ischemia/reperfusion, and the number was increased gradually and deeply stained as the time of reperfusion prolonged (Figure 3), it reached the peak value at 1-3 days, and then weakened gradually, but it was still higher than in the sham-operated group at 14 days [(5.01 ±1.71), (5.21 ± 1.62) cells/visual field; (2.03± 1.73),(2.46± 1.38) cells/visual field, P ＜ 0.05]. ④ In the sham-operated group, lightly stained FGFR-1 positive cells could be observed in brain tissue. At 6 hours after cerebral ischemia/reperfusion, the FGFR positive cells began to increased in cortexes and striatums, which were the most at 1-3 days, and gradually decreased after 3 days, and the number was still a little more than that in the sham-operated group at 14 days [(5.01± 1.41), (5.20± 1.33) cells/visual field; (2.25±1.67),(2.32± 1.61 ) cells/visual field].CONCLUSION: After cerebral ischemia/reperfusion, the expressions of endogenous bFGF and FGFR-1 may be activated in cortex and striatum, then inhibit the neuronal apoptosis, and play its neuroprotective role.

Objective To investigate the prognostic factors and survival of patients with bronchopulmonary carcinoid tumors (BPC) after surgical treatment .Methods The clinical data of 87 patients undergoing surgery for BPC from Jan .2002 to Dec .2008 were reviewed retrospectively .Kaplan-Meier method was used to analyze the survival of the patients .The risk factors such as age , gender ,smoking history ,histological type ,tumor size ,were analyzed by Cox proportional hazards regression model .Results The 1-,3-and 5-year overall survival rates were 85 .1% ,71 .3% and 63 .2% .Univariate analysis revealed that age (P=0 .016) ,smoking history(P=0 .007) ,histological type(P=0 .000) ,tumor stage(P= 0 .000) ,tumor size(P= 0 .006) lymph node metastasis(P=0 .000) ,surgery type (P= 0 .045) and postoperative chemotherapy (P= 0 .000) were prognostic factors .Multivariate analysis showed that histological type(P=0 .008) ,tumor stage(P=0 .000) ,lymph node metastasis(P=0 .033) were independent prognostic factor .Conclusion The survival rate of the BPC patient after surgical treatment is high ,histological type ,tumor stage and lymph node metastasis were independent prognostic factors .

Objective To investigate the neuroprotective effects and mechanism of phycocyanin in cerebral ischemia reperfusion injury in rats.Methods The model of middle cerebral artery occlusion reperfusion(MCAO/R) was established using the intraluminal filament occlusion with healthy adult male Wistar rats treated by phycocyanin.The apoptosis and the expression of bFGF and FGFR-1,IGF-1 and IGF-1R,iNOS and SOD were respectively determined by TUNEL and immunohistochemical staining to evaluate the effects of phycocyanin on above indexes.Results ① The rats showed neurobehavioral function disorders and the number of nerve cells reduced while apoptotic cells increased in ischemic area after ischemic reperfusion.In phycocyanin group,the number of apoptotic cells reduced siginificantly during reperfusion 12h~3d and the neurobehavioral function was better than that those in control group during reperfusion 7~14d.② In control group,the expressions of bFGF and FGFR-1 increased successfully from reperfusion 6h and reached a maximum at 1d,then subsided gradually in cortex and striatum.In phycocyanin group,the numbers of bFGF and FGFR-1 positive cells were higher than those in control group at the same time-points,which were significantly at reperfusion 1d and 12h~3d respectively.③ The expressions of IGF-1 and IGF-1R increased in cortex and striatum following cerebral ischemic and reperfusion.In phycocyanin group,the numbers of IGF-1 and IGF-1R positive cells in each time-point were higher than those in control group,which was significantly during reperfusion 6h~1d.④ In cotex and striatum,the iNOS and SOD expressed strongly and keep high level during 6h~7d with the maximum at reperfusion 1d.In phycocyanin group,iNOS expressed significantly higher during 12h~1d whlie SOD lower during 6h~1d than those in control group at the same time-point.Conclusion Phycocyanin might play a intrinsic antioxide effect by up-regulating SOD and down-regulating iNOS to inhibit neuronal apoptosis,and enhance the neuronal repairation by means of inducing the expressions of bFGF and IGF-1 following cerebral ischemic reperfusion in rats.

To summarize the present situation of research on amylolytic lactic acid bacteria and its amylase,and the fermentation technology of amylolytic lactic acid bacteria.The amylolytic lactic acid bacteria have the ability of secreting amylase,can direct ferment starchy material into lactic acid without hydrolysising processes,simplify the production process,and saving the equipment investment,thereby,decrease the cost of production.Amylolytic lactic acid bacteria isolated from traditional fermented food,or from organic waste and kitchen waste.The mechanism of direct utilize starch raw materials by the amylolytic lactic acid bacteria were introduced,the fermentation technology of amylolytic lactic acid bacteria were compared.Meanwhile it was pointed out that through mutation breeding and genetic engineering methods gain the potent amylolytic lactic acid bacteria,and use the advanced fermentation and separation technology can improve the productivity of lactic acid.

Objective To explore the effects of a single bout of aerobic exercise on oxidative stress and total peripheral vascular resistance. Methods 36 prehypertensive patients were divided into aerobic exercise group (n=18) and control group (n=18). The aerobic exercise group performed cycle ergometry at 70% maximal oxygen consumption for 30 min. Total peripheral vascular resistance (TPVR), malondialdehyde (MDA), mean arterial pressure (MAP), vitamin C (VC) and vitamin E (VE) were measured before, immediately, 1 hour and 2 hours after the aerobic exercise. Results There was no significant difference in TPVR, MDA, MAP, VC and VE between two groups before exercise (P>0.05). TPVR was all significantly lower in the aerobic exercise group than in the control group immediately, 1 hour and 2 hours after exercise (P<0.001), and showed a rising trend of recovery. MDA was significantly higher in the aerobic exercise group than in the control group 1 hour and 2 hours after exercise (P<0.05), however there was no significant difference in MDA between two groups immediately after exercise (P>0.05). MAP was significantly lower immediately after exercise in the aerobic exercise group than in the control group (P< 0.001), but there was no significant 1 hour and 2 hours after exercise (P>0.05). There was no significant difference in VC and VE between two groups immediately, 1 hour and 2 hours after exercise (P>0.05). Conclusion The level of oxidative stress increases after a single bout of aerobic exercise, and the TPVR decreases.

Objective To investigate the effect of direct intercell contact on the bone mesenchymal stem cells(MSCs)differentiate into nucleus pulposus cells(NPs)when cocultured with NPs in constant magnetic field.Methods The primary NPs labeled by DAP1 were cocultured with the 3 rd generation of MSCs through direct and indirect intercell contact in the presence or absence of constant magnetic field(0.05,0.10,0.50 and 1.00 mT,respectively). Observation of morphological changes was performed every 24 hours.The method of MTT was employed to evaluate the level of proliferation.The gene expression of collagen Ⅱ,Sox-9 and Aggrecan was measured by using RT-PCR. Results MSCs cocultured with direct intercell contact with the NPs rounded up and presented a round ring-like structure appearance.The expression of marker genes including Collagen type Ⅱ,Aggrecan and Sox-9 were significantly increased when cells cocultured in constant magnetic field of 0.05 mT compared with those without constant magnetic field(P＜0.05).There were no significantly changes with regard to the expression of the above genes in 0.10 mT field(P＞0.05).The growth of NP-like cells was suppressed when the intensity of magnetic field was higher than 0.10 mT(P＜0.05).Conclusion It is suggested that 0.05 mT constant magnetic field and direct intercell contact facilitate differentiation of MSCs into NPs.

To offer the reference and method for salt damage in the cultivation of Marsdenia tenacissima, the seeds of M. tenacissima collected from Maguan city ( Yunnan province) were taken as the test materials to study the effects of different priming materials on improving germination and growth under high-level salt stress condition. Four different treatments, which were GA3, KNO3-KH2PO4, PEG-6000, NaCl, combined with ANOVA were applied to test the performance of germination energy, germination percentage, germination index, MDA, SOD, and CAT. The results showed that the seed germination was obviously inhibited under salt stress and the soaked seeds with different priming materials could alleviate the damage of salt stress. Under these treatments, the activities of SOD, CAT the content of soluble protein significantly increased. While the content of MDA significantly decreased. The maximum index was obtained when treated with 1.20% KNO3-KH2PO4, the germination percentage increased from 52.67% to 87.33% and the activity of SOD increased from 138.01 to 219.44 respectively. Comparing with the treatment of 1.20% KNO3-KH2PO4, the germination percentage of treating with 300 mg x L(-1) GA3 increased from 52.67% to 80.67%, while the activity of SOD increased from 138.01 to 444.61.
Germination ; drug effects ; physiology ; Marsdenia ; drug effects ; growth & development ; Nitrates ; pharmacology ; Polyethylene Glycols ; pharmacology ; Potassium Compounds ; pharmacology ; Seeds ; drug effects ; growth & development ; Sodium Chloride ; pharmacology ; Stress, Physiological ; Xanthones ; pharmacology

In this paper, Fourier transform infrared spectroscopy fingerprint analysis of Marsdenia tenacissima samples was used to develop a reliable method of tracing the geographical origins. Forty-eight samples from four provinces of China were analyzed by FTIR. We analyzed and characterized the fingerprints in both the full spectrum peaks and characteristic peaks, then the principal component analysis and the cluster analysis were carried out. The results of fingerprint analysis, correlation analysis, principal component analysis and cluster analysis can identify the geographic origins correctly, which verified and supplemented each other; the identification results and the actual location showed a high degree of consistency, namely the lower the space distance, the greater the similarity of different samples. These results revealed the obvious superiority and practical value in comparison to the more tedious and time-consuming wet chemistry method normally used. Using appropriate metrology methods can trace the geographical source correctly. The M. tenacissima materials from the region of Maguan should be considered as genuine medicinal materials taking into account the good quality.
China ; Cluster Analysis ; Drugs, Chinese Herbal ; analysis ; classification ; standards ; Geography ; Marsdenia ; chemistry ; classification ; Medicine, Chinese Traditional ; Principal Component Analysis ; Quality Control ; Reproducibility of Results ; Spectroscopy, Fourier Transform Infrared ; methods

Accidents, Occupational ; statistics & numerical data ; Adult ; Extraction and Processing Industry ; Female ; Humans ; Male ; Petroleum