Objective To explore the effect of protocatechuic acid on serum tumor necrosis factor-α( TNF-α) , interleukin-1β( IL-1β) and oxidative stress products levels in Parkinson rats.Methods 60 male SD rats were randomly divided into normal control group ( n=10 ) , model group (n=10), madopar group (n=20) and protocatechuic acid group (n=20).Rat model with Parkinson disease were builded in model group, madopar group and protocatechuic acid group.Madopar group and protocatechuic acid group were given corresponding drug with a consecutive treatment of two weeks.After treatment,the serum TNF-α,malondialdehyde (MDA), superoxide dismutase (SOD) and IL-1βlevels were detected in all groups.Results Compared with normal control group, the serum TNF-α, IL-1βand MDA levels in model group were significantly higher, and SOD level was lower (P<0.05).Compared with model group, the serum TNF-α, IL-1βand MDA levels in madopar group pre-treatment were significantly lower, and SOD level was higher (P<0.05).There were no significant difference of serum TNF-α, IL-1β, MDA and SOD levels between madopar group and protocatechuic acid group.Conclusion The protocatechuic acid could significantly reduce the serum TNF-α, MDA and IL-1βlevels in Parkinson model rats, enhance the activity of SOD, which has protective effect on oxidative stress injury induced by Parkinson disease.

Objective To investigate the effectiveness and safety of endovascular coiling and microsurgical clipping for ruptured intracranial aneurysms. Methods Patients w ith ruptured intracranial aneurysm treated w ith endovascular coiling or microsurgical clipping w ere enrol ed retrospectively. The demography, baseline clinical data, outcome, and complications in patients received endovascular coiling and microsurgical clipping w ere compared. Results A total of 85 patients w ith ruptured intracranial aneurysm were enroled, including 40 were treated with microsurgical clipping (surgical clipping group) and 45 were treated w ith endovascular coiling (endovascular coiling group). There w ere no significant differences in the proportions of the patients in male (37.5%vs.40.0%; χ2 =0.056, P=0.813), hypertension (30.0%vs. 33.3%; χ2 =0.109, P=0.742 ), smoking ( 50.0%vs.48.9%; χ2 =0.010, P=0.918 ), drinking (45.0%vs.46.7%; χ2 =0.024, P=0.878), aneurysm site (anterior communicating artery: 50.0%vs. 48.9%;posterior communicating artery:35.0%vs.33.3%; middle cerebral artery:10.0 %vs.11.1%;vertebral artery: 5.0%vs.6.7%; al P>0.05), aneurysm maximum diameter < 10 mm (80.0%vs. 77.8%;χ2 =0.063, P=0.802), Hunt-Hess grade 1-2 (55.0%vs.57.8%; χ2 =0.066, P=0.797), Fisher grade 1-2 ( 60.0%vs.57.8%; χ2 =0.043, P=0.835 ), and time from onset to treatment < 72 h (62.5%vs.64.4%; χ2 =0.035, P=0.853) in the surgical clipping group and endovascular coiling group. There w ere no significant differences in the complete occlusion rate of aneurysms ( 97.5%vs.91.1%;P=0.364) and the good outcome rate (65.0%vs.68.9%; χ2 =0.145, P=0.703) betw een the surgical clipping group and the endovascular coiling group. No patients died in the surgical clipping group and 1 patient died in the endovascular coiling group, and there w as no significant difference ( P=1.000). One patient (2.5%) had cerebral infarction in the surgical clipping group and no patients had cerebral infarction in the endovascular coiling group, and there w as no significant difference ( P=0.471). Conclusions The efficacy and safety of microsurgical clipping are the same as those of endovascular coiling for ruptured intracranial aneurysms.

Objective To compare the efficacy, complications, safety and prognosis of the minimally invasive puncture approach and key hole in the treatment of hypertensive cerebral hematoma.Methods A totol of 68 patients with hypertensive cerebral hematoma confirmed by CT from April 2012 to October 2013 in Nongken Sanya Hospital were randomly divided into key hole evacuation group(n=32) and minimally invasive puncture group (n =36).Comparisons were made between the two surgical methods in the operative time, postoperative complications, the fatality and the postoperative re-haemorrhagia rate, neurological function deficit score also been observed and evaluated in the 1 st,2nd and 4th weeks after surgery.Results The NFDS scores of the two groups both decreased in the 1st week after surgery,but compare with preoperative the difference was not statistically significant (P ＞ 0.05).In the 2nd weeks and 4th weeks after surgery, NFDS scores further decreased in both group,and there was statistically significant compare with preoperative(the key hole evacuation group : (26.2±4.5) vs.(17.8 ± 3.6) vs.(44.1 ± 5.4) scores;the minimally invasive puncture group: (22.1 ± ±3.7) vs.(15.4±2.8) vs.(43.9±6.2)scores;P＜0.05) ,but during the same period there was no significant difference between the two groups with NFDS scores(P＞0.05).The rebleeding rate of the minimally invasive puncture group was significantly lower than the key hole evacuation group (4.08％ vs.16.33％, x2=6.56, P＜0.05).There was no significant difference in mortality rate and long term total effect between two groups (P＞0.05).Conclusion Although both key hole and minimally invasive puncture are effective measures for treatment of hypertensive cerebral hemorrhage, but minimally invasive puncture with less trauma, definite curative effect and higher security advantages in clinical.

BACKGROUND:Animal models of osteoporosis play an important role in the research of the pathogenesis, occurrence and development of osteoporosis, as wel as in the clinical diagnosis, prevention and treatment of osteoporosis. OBJECTIVE: To summarize and discuss the establishment and research ideas of osteoporosis models, explore the current situation and advance of osteoporosis models, compare the advantages and disadvantages of various methods, and provide evidence for clinical investigation. METHODS: A computer-based online search was conducted in SinoMed, VIP, Wanfang and PubMed databases by using the key words of “animal model, osteoporosis” from January 1969 to October 2015. The language was limited to both Chinese and English. Relevant articles were screened according to inclusion and exclusion criteria. The documents about the methods of osteoporosis model preparation, method improvement as wel as their advantage and disadvantage were summarized. RESULTS AND CONCLUSION:A total of 576 articles were included. Among them, articles published earlier, duplicated, and similarly were excluded, and 53 articles were finaly included. Various animal models of osteoporosis may only focus on the certain causes, certain stage, some of the main symptoms and some pathophysiological changes of disease. Accordingly, appropriate modeling methods and experimental animals should be selected based on research objective. Rat undergoing castration is the most commonly used model in the modeling of osteoporosis. Among drug methods for constructing osteoporosis model, glucocorticoids is the most commonly used one. Disuse method and nutritional method have limitations, and always combined with castration and drug methods. The effects of gene transfer, gene mutation and brain-derived model deserve

Objective:To observe the up-regulation of nuclear Clusterin (nCLU)gene on the biological behaviors of human non-small cell lung cancer cell line A549 .Methods:Sense eukaryotic expression vector of nCLU was constructed by cloning the cDNA of nCLU into pIREShyg3 vector. A549 cells were transfected with pIRES-nCLU and pIREShyg3 vectors by lipofectAMINE~(TM) 2000 mediation, respectively. Stable transfected cells were selected by hygromycin B screening. CCK-8 assay was performed to evaluate the effect of nCLU over-expression on cell proliferation in vitro. The expression level of nGLU protein was examined by Western blotting. Cell cycle distribution was detected by FCM with PI staining. The alteration of migration and metastasis potential of A549 cells before and after nCLU gene transfection were assayed by cell chemotactic migration and invasion test. Results:The proliferation speed of the transfected A549 cell clones stably over-expressing nCLU was slowed down. FCM analyses revealed that the percentage of cells in G_0/G_1 phase dramatically increased from (33.54±2.10)% to (63.31±4.30)%. The cell chemotactic migration and invasion potentials were markedly reduced after nCLU gene transfection (P<0.05). Conclusion:Up-regulation of nCLU can greatly inhibited the proliferation and decreased the migration and invasion capabilities of A549 cells.

Objective To observe the effect of pituitary adenylate cyclase activiting polypeptide (PACAP) on traumatic brain injury (TBI) and on tbeCD4+/CD8+ T cell number in blood and spleen of rats.MethodsThe male SD rats were randomly (random number) divided into sham operation group ( n =6),normal saline + TBI group ( n =6) and PACAP + TBI group ( n =6).Right parietal cortical contusion was produced by a weight-dropping method.PACAP was administered intra-cerebroventricularly in a dose of 1 μg/5 μl 20 min before TBI.Brain tissue samples were taken 24 h after trauma.The injured brain tissue of rats was examined by HE stain.The numbers of CD4+/CD8+ T cells in blood and spleen were deteced with flow cytometry.Results Edema,hemorrhage,inflammatory cell infiltration,swollen,degenerated neurons and neurons arrayed disorderly around the injured cortex in hippocampus were found under light microscope.The average numbers of CD4 + T lymphocytes counts in blood and spleen were lower ( P =0.000,P =0.005 ),and number of CD8 + T lymphocytes was higher ( P =0.01 ) in TBI rats group than those in the sham operation group.Micro-injection of PACAP into cerebroventricular attenuated the injury,significantly increased the number of CD4 + T lymphocytes in blood and spleen ( P =0.019,P =0.839),and decreased the number of CD8 + T lymphocytes.ConclusionsPretreatment with PACAP may protect against TBI via influencing periphery T cellular immune function.

Objective:To investigate the anti-tumor effect of Hyperoside.Methods: Human γδT cells were amplified by isopentenyl pyrophosphate from peripheral blood cells.The proliferation capacity of γδT cells was measured with CCK-8 assay after treated with different concentrations of Hyperoside.Cytotoxicity of γδT cells was detected with LDH assay , and the expression of granzyme,perforin CD107a and IFN-γonγδT cells were measured by flow cytometry before and after treatment.Results: Hyperoside could significantly stimulate the proliferation of γδT cells at the concentration of 3.13-12.5 μg/ml.Cytotoxicity and expression of granzyme,perforin and IFN-γofγδT cells were increased after treatment.Conclusion:Hyperoside could enhance cytotoxicity of humanγδT cells through up-regulation of granzyme ,perforin CD107 a and IFN-γexpression.

Objective To investigate the in vitro effects of quercitrin on the proliferation and the cytotoxicity of human γδT cells.Methods Peripheral blood mononuclear cells (PBMCs) were isolated from healthy subjects and cultured with isopentenyl pyrophosphate and IL -2 to induce human γδT cells.The hu-manγδT cells were cultured with quercitrin at various concentrations for 48 hours.CCK-8 kits were used to analyze the in vitro proliferation and cytotoxic activities of γδT cells.Flow cytometry was performed to meas-ure the expression of granzyme B and perforin in γδT cells.The expression of p-ERK, p-Akt and Bcl-2 at protein level were detected by Western blot .Results The percentage of human γδT cells in PBMCs was in-creased from (2.96±1.83)%to (88.94±2.36)%after 10 days of culture.The quercitrin at concentrations of 10 to 80 μg/ml could promote the growth of γδT cells and up-regulate the expression of granzyme B , per-forin, p-ERK, p-Akt and Bcl-2 in a dose dependent manner .The cytolytic activities of γδT cells against co-lonic carcinoma cells ( HCT116 ) were enhanced by quercitrin .Conclusion Quercitrin could promote the proliferation of γδT cells and enhance the expression of granzyme B and perforin at certain concentrations in vitro.ERK1/2 and Akt signal transduction systems might be involved in the process .

ObjectiveTo observe the effect of betulinic acid(BetA) on the growth of human cytokine induced killer(CIK) cells and the killing activity of CIK cells on the gastric cancer cells in vitro before and after induced by betulinic acid,explore its mechanism.MethodsPeripheral blood mononuclear cell (PBMC) were separated form the healthy and were induced with various of cytokine to become CIK cells in vitro.CIK cells were collected on the tenth day and were induced with betulinic acid in different concentrations,followed by 48 h,the colorimetric methyl thiazolyl tetrazolium(MTT) method assay the proliferation rate of human CIK cells.Flow cytometry (FCM) was used to detect the expression changes of perforin,granzyme B and CD107a of human CIK cells before and after betulinic acid-induced.Lactate dehydrogenase (LDH) release assay was used to measure the influence on cytotoxic activity of CIK cells induced by betulinic acid against gastric cancer cell line SGC-7901 in vitro.Western blot assay was used to measure the extracellular signal-regulated kinase1/2 (ERK1/2),and adapter proteins SH2-domain containing leukocyte protein of 76KD(SLP-76) and linker for activative of T cells(LAT) expression changes of human CIK cells before and after drug-induced.ResultsBetulinic acid can promote CIK cells growth when the concentration were in 0.08-10 μg/ml,the expression of perforin,granzyme B and CD107a of CIK cells were significantly higher than control group(P＜0.05) when the concentration of betulinic acid were in 0.3 μg/ml.In the meanwhile,the cytotoxic activity of CIK cells in vitro against gastric cancer cell line SGC-7901 were also remarkably higher than the control group (P＜0.05).The expression of SLP-76,LAT and ERK1/2 were significantly increased to a certain extent than the control group( P＜0.05 ),when CIK cells were treated with betulinic acid.ConclusionThese results suggest that betulinic acid can promote CIK cells growth in some concentrations and increase the cytotoxic activity of CIK cells against gastric cancer cell line SGC-7901,its mechanism may related with two factors,on the one hand,enhancing the activity of SLP-76,LAT and ERK1/2,on the other hand,increasing the expression of perforin,granzyme B and CD107a on the surface of CIK cells.

Objective To study the relationship between the cognitive function and speech recognition ability in young patients with OSAHS.Methods We selected 60 young male patients,according to the apnea-hypopnea index(AHI)and the severity of hypoxemia.They were divided into three subgroups on the basis of their syndrome severities:mild group (n= 19;AHI 5~15/h,85%≤minimum SaO2≤90%),moderate group (n= 20;AHI>15~30/h,80%≤minimum SaO2<85%),and severe group (n= 21;AHI>30/h,minimum SaO2<80%).First,we used the MoCA scale for cognitive function tests and recorded the scores.Then 15 lists of sentence Mandarin Speech Test Materials(MSTMs)were utilized to test each group.A data analysis was performed using SPSS 17.0 software. Results The total MoCA scores(mild group:27.32±1.16;moderate group:25.85±1.23;severe group:24.52± 1.69;control group:28.52 ±1.16)decreased progressively as the disease severity increased,showing significant differences between the control group and the mild,moderate and severe groups of OSAHS patients (allP<0.05). When sound stimuli were presented at 22,24,and 26 dB SPL,the speech recognition rates in the patients with se-vere(35.4±22.6,56.3±23.9,75.2±16.5)lower than the other groups (mild group:38.4±23.5,58.3±25.5,79.2 ±18.5;moderate group:38.8±21.6,58.7±22.7,78.5±16.7;control group:39.4±23.5,60.3±24.3,80.2±16.4, respectively,allP<0.05).The differences in intensity of 50% recognition rate between the severe group(4.15± 0.80)and the control(3.62±0.41),mild (3.66±0.50)and moderate groups(3.72±0.55)of OSAHS patients were statistically significant(allP<0.05).Conclusion With hypoxia and disease severity increased,speech recogni-tion abilities in OSAHS patients decreased.This may be an important factor associated with cognitive assessment scale score.