Objective To study intervention effect of diet habits according to Trans-theoretic Model (TTM) in young and middle-aged patients with asymptomatic hyperuricemia. Methods 120 patients were divided into the intervention group (n=60) and the control group (n=60). The nursing intervention by stages of TTM was implemented to a total of 56 patients in the intervention group in the study. Routine nursing care was taken to 57 patients in the control group. Patients were evaluated by self-designed questionnaire after 1, 3 and 6 months of the intervention in the two groups respectively, including the diets of patients, the change of patients′ eating habit, differences of uric acid, total cholesterol, triglycerides, blood sugar. Results The patients′ intake of beer, seafood, meat, edible oil in the intervention group was lower than the patients′ intake in the control group, while the patients′ intake of fruits and vegetables was higher than the patients′intake in the control group after 1, 3 and 6 months of the intervention, which was statistically significant ( P < 0.01). Compared with 27 cases in the control group, 53 cases in the intervention group were in action phase and maintenance phase after 6-month intervention, the difference was statistically significant (P < 0.01). After six months′intervention, the patients′uric acid, triglycerides in the intervention group was (418.01 ± 24.15)μmol/L and (1.52 ± 0.56) mmol/ L respectively, which was significantly lower than (435.02 ± 26.35) μmol/ L and (2.45 ± 0.74) mmol/L in the control group, t values were-3.47 and 2.20 respectively, P<0.01. Conclusions Diet intervention based on the TTM can effectively promote the change of patients′ unhealthy eating habits, improve self-management skills, and reduce uric acid, which is worthy of generalizing.

Objective To investigate the neuralprotective effect of Rho kinase inhibitor fasudil hydrochloride in cerebral ischemia/reperfusion injury in rats.Methods The SD rats were randomly divided into four groups:the sham group,the ischemia/reperfusion group,the fasudil hydrochloride group and the physiological saline group.Fasudil hydrochloride were injected intraperitoneally 30 minutes before ischemia.And the physiological saline group were treated with the intraperitoneal injection of the same volume of saline.The phosphorylation and protein expression of GluR6 at 6 hours during reperfusion were detected using immunoprecipitation and immunoblotting analysis to examine the effect of Fasudil hydrochloride.Furthermore,TUNEL staining was used to examine the apoptosis of neurons in rat hippocampal CA1 regions after 3 days reperfusion.Results 1.Immunoprecipitation and immunoblotting analysis were used to analyze the phosphorylation of GluR6 in serine site.The results showed that the GluR6 serine phosphorylation level increased significantly at 6h of reperfusion compared with the sham group (P＜0.05).Fasudil hydrochloride group could inhibit the increased phosphorylation of GluR6 at 6h of reperfusion compared with the ischemia/reperfusion group and saline group,respectively (P ＜ 0.05).2.TUNEL staining was used to examine the apoptosis of neurons in 3 days after reperfusion in CA1 regions of hippocampus.The results indicated that significant numbers of TUNEL positive cells (40.20 ± 2.77) were observed 3 days after ischemia/reperfusion.The numbers of viable neurons per 1 mm length of CA1 pyramidal cells were quantitatively analyzed.Fasudil hydrochloride markedly decreased the neuronal loss compared with the ischemia/reperfusion group (19.80 ± 2.86) (P＜0.05).Conclusion Fasudil hydrochloride can inhibit induced phosphorylation of GluR6 by the ischemia/reperfusion.Fasudil hydrochloride can reduce the neurons apoptosis in hippocampal CA1 regions,and perform a neuralprotective effect on ischemia/reperfusion injury in rats.

Objective To investigate the effects of the FKBP51 · PHLPP · AKT signal module on the phosphorylation of Akt and hippocampal neuronal injury after the cerebral ischemia / reperfusion induced neuronal death in rat hippocampus.Methods Transient(15 min)brain ischemia was induced by the four-vessel occlusion in Sprague-Dawley rats.6 rats were used in each group.The antisense oligodeoxynucletides(AS ODN)of PHLPP2 (PH domain and leucine rich repeat protein phosphatases) was used to suppress the assembly of FKBP51 · PHLPP · Akt signal module by intracerebroventricular infusion once per day for 3 days before ischemia.After 6 hours reperfusion,interactions of PHLPP2 and FKBP51 (FK506 binding protein 5) with Akt were detected by immunoprecipitation (IP) and the phosphorylation of Akt was detected by western blot (IB).After 5 days reperfusion,rats were perfusion-fixed with paraformaldehyde and Hematoxylin-Eosin staining was used to examine the survival number of CA1 pyramidal cells of hippocampus.Results Compared to PHLPP2 MS ODN group(1.24±0.24,1.68±0.11,0.58±0.01),PHLPP2 AS ODN suppressed the assembly of the FKBP51 · PHLPP · Akt signaling module(1.06±0.01,1.04±0.13),and increased the phosphorylation of Akt(0.76±0.02) (P＜0.05).Furthermore,compared to PHLPP2 MS ODN group (20.1±2.5),the number of surviving neurons significantly increased in PHLPP2 AS ODN group(88.3±2.7)(P＜0.05).Conclusion The increasing assembly of FKBP51 · PHLPP · Akt signal module can damage CA1 pyramidal cells of hippocampus by inhibiting the phosphorylation level of Akt.

Objective To investigate the effect of Rho kinase inhibitor fasudil on taxed lineage kinase 3 (MLK3), c-Jun NH2-terminal kinase (JNK) phosphorylation, caspase-3expression, and neuronal injury in hippocampal CA1 region follwong cerebral ischemic rep erfusion in rats. Methods A total of 72 Sprague-Dawley rats were randomly divided into sham operation, ischemia-reperfusion, normal saline, and fasudil groups. A global cerebral ischemic model was prepared by four-vessel ligation. The levels of MLK3 and JNK phosphorylation, and caspase-3 expression were detected by Western blot analysis. Cresy1 violet staining was used to detect the numbers of survival neurons in hippocampal CA1 region. Results When 6 hours after ischemia-reperfusion, the level of MLK3 phosphorylation in the fasudil group (1.13 ± 0. 03)was significantly lower than that in the normal saline group (2. 08 ± 0. 01 ,P = 0. 000 3), while the levels of MLK3 was no significant difference. When 3 hours after ischemia-reperfusion, the level of JNK phosphorylation in the fasudil group (1.27 ±0. 02)was significantly lower than that in the normal saline group (2.09 ±0. 01, P=0. 000 2), while the levels of JNK was no significant difference. When 6 hours after ischemia-reperfusion, the expression level of caspase-3in the fasudil group (1.28 ± 0. 02) was significantly lower than that in the normal saline group (2. 10 ± 0. 01, P = 0. 000 6). When 5 days after ischemia-reperfusion, the pyramidal cells in hippocampal CA1 region almost completely disappeared in the ischemia-reperfusion group, and only a few cells left (9. 8 ±2. 1). The numbers of survival pyramidal cell (8. 28 ± 3.2) in hippocampal CA1 region in the fasudil group was significantly more than that in the normal saline group (11.8 ± 1.6, P ＜0. 05). Conclusions Fasudil may significantly inhibit the ischemia-reperfusion-induced phosphorylation of MLK3 and JNK, as well as the expression of caspase-3, and thus reduce neuronal injury in hippocampal CA1 region.

Objective: To analyze the influence of semen processing methods on the proportion of the aneuploid sperm by detecting the sperm's chromosome X, Y, 18 using fluorescence in situ hybridization. Methods: Ten patients with mild ohgoasthenosperia, who were received ICSI treatment, were included in this study. Five semen samples of the patients were randomly selected to detect using Swim-up method (A group) and 5 using sperm-grad double-density centrifugation method (B group).Another 5 patients with mild oligo-asthenosperia were as control (C group). The CEPX / Y and CEP18 probe was used to detect the sperm of these 15 patients by fluorescence in situ hybridization. The proportion of aneuploid sperm was compared in three groups. Results: The sex chromosome aneuploid rates were (4.21±2.49)%, (3.24～1.49) % and (2.62±0.89) % in control, A and B groups. The rates of aneuploid chromosome 18 were (3.00±1.22)%, (2.00～1.22)% and (2.00±1.22)% in control, A and B groups. There were no significant differences in three groups (P＞0.05). Conclusion: The results showed that the methods of Swim-up and Sperm-Grad double-density gradient centrifugation could select sperms in motility potential and teratospermia,but not in normal chromosome sperms.

Objective To investigate effect of FK506 binding protein 51 (FKBP51) on the c-JunN-terminal kinase (JNK) pathway in cerebral ischemia-reperfusion injury.Methods Transient global cerebral ischemia rat models were made by four-vessel method.Healthy male SD (Sprague Dawley) rats were randomly divided into:sham group,ischemia/reperfusion group (I/R group),FKBP51 antisense oligonucleotide group (FKBP51 ASODN group),FKBP51 missense oligonucleotide group (FKBP51 MSODN group),and solvent control group (TE group).The effect of FKBP51 ASODN on expression of FKBP51 protein and JNK was detected,and c-Jun phosphorylation was detected by Western blot.Results (1) FKBP51 protein expression in the FKBP51 ASODN group was reduced.The change of FKBP51 protein expression between the FKBP51 ASODN and sham groups was statistically significant (P ＜ 0.05).(2) The expression differences of total JNK protein between all the groups were not statistically significant (P ＞ 0.05).The expression of p-JNK in sham group was significantly less than the other groups (P ＜ 0.05).The expressions of p-JNK in I/R 3d,TE,and FKBP51 MSODN groups were higher relative to Sham group; however,the differences among those three groups were not statistically significant (P ＞ 0.05).The expression of p-JNK in FKBP51 ASODN group was significantly less than FKBP51 MSODN group (P ＜ 0.05).(3) The expression differences of total c-Jun protein among all groups were not statistically significant (P ＞ 0.05).The expression of p-c-Jun in sham group was significantly less than the other groups (P ＜ O.05).The expressions of p-c-Jun in I/R 6 h,TE,and FKBP51 MSODN groups were higher relative to the sham group; however,the differences among those three groups were not statistically significant (P ＞ 0.05).The expressions of p-c-Jun in FKBP51 ASODN group was significantly less than FKBP51 MSODN group (P ＜ 0.05).Conclusions FKBP51 might activate JNK signaling pathway in cerebral ischemia-reperfusion injury.

Objective To investigate the biological effects of bcl-2 gene on neurons. Methods The recombinant expression plasmid pc DNA3-bcl-2 was constructed from pSFFV-bcl-2,then it was introduced into PC12 cell line by liposome method.Western blotting and immunohistochemistry in situ were applied to exam the exogenous gene expression.The two groups of cells(Group A,PC12 transfected by pcDNA3-bcl-2 and Group B,PC12 transfected by pcDNA3) were exposed to cisplatin with the concentration of 10*!?mol/L,50*!?mol/L, and 100*!?mol/L 72 hours later,the survival cells were estimated.Cell cycle indexes between these two groups were also studied by FCM. Results The recombinant expression plasmid pcDNA 3-bcl-2 was constructed successfully and PC12 cell line transfected by the plasmid could express Bcl-2 protein effectively.Compared with the control(25 79%),there was a significant decrease of cells from the S phase in PC12 with bcl-2 gene(8.81%).After exposed with 10*!?mol/L,50*!?mol/L,and 100*!?mol/L cisplatin,the surviving cells in group A were 276?13,185?11 and 108?10 respectively,which increased much more than in group B while they were 100?9,12?3 and 2?2 accordingly.Conclusions bcl-2 can protect PC12 cells against cytotoxic insults of cisplatin,and it suggested that it might act via cell cycle controlling.

Objective To explore the effect of FKBP51 acting on Caspase-3 and hippocampal CA1 area neu-ronal necrosis in cerebral ischemia reperfusion injury of rat.Methods SD rats were randomly divided into Sham group,ischemia reperfusion group ( I/R group ) , TE buffer group ( TE group ) , FKBP51 antisense oligonucleotide group (FKBP51 ASODN group) and FKBP51 missense oligonucleotide group (FKBP51 MSODN group).Transient global cerebral ischemia rats models were made by four-vessel method.We used Western blot to detect the expression of FKBP51,the effect of FKBP51 ASODN to FKBP51 expression and Caspase-3 activity;while we used HE staining technique to detect FKBP51 ASODN effect to rat hippocampal CA1 area neuronal necrosis.Results (1) In Sham group and I/R group (0min,15min,30min,1h,3h,6h,1d,3d),FKBP51 expressed,and the difference among the groups was no statistical significance (F=0.64,P>0.05).(2)The expression of FKBP51 in FKBP51 ASODN group was obviously reduced, and the difference was statistically significant compared with Sham group ( t =8.21, P <0.05).(3)The expression of Cleaved-Caspase-3 in Sham group obviously declined than the other groups,the differ-ence between them was statistically significant (F=12.31,P<0.05);The expression of FKBP51 in FKBP51 ASODN group was decreasing compared with FKBP51 MSODN group,and the difference was statistical significance(t=9.71, P<0.05).(4)HE staining showed:the number of Sham group (186.3 ±2.5) hippocampal CA1 pyramidal cells was most.The cells arranged densely,and nucleoli were large and round,the difference was statistically significant com-pared with the other groups (χ2 =81.91,P<0.05);The hippocampal CA1 pyramidal cells of I/R group (15.4 ± 2.6),TE group (18.5 ±2.2) and FKBP51 MSODN group (17.5 ±1.8) were almost completely disappeared,only left a few residual cells,a great quantity of denaturated cells which presented karyopykosis,tinctorialed endochylema, ruptured of membrane and released cell content;the hippocampal CA1 pyramidal cells FKBP51 ASODN group (92.8 ±2.6) survival increased significantly compared with other group,the difference was statistically significant (χ2 =52.36,P<0.05).Conclusion In cerebral ischemia reperfusion injury,FKBP51 can enhance the activation of Caspase-3 (Cleaved-Caspase-3) expression and inhibit the survival of the neurons.

Objective To investigate the effects of subcutaneous transfection of human beta-nerve growth factor (Ad-hNGFβ) gene on the expression of substance P (SP) in the dorsal root ganglion in a rat model of diabetic neuropathic pain (DNP).Methods Male SD rats weighing 180-220 g were used in this study.Ten rats were randomly collected as normal control without DNP (group C).DNP model was made by intraperitoneal injection of streptozocin (STZ) 75 mg/kg.Seventy-five rats with DNP were randomly divided into 3 groups ( n =25 each):DNP group; Ad-hNGF group and Ad-LacZ group.Groups Ad-NGF and Ad-LacZ received subcutaneous transfection of 1.12 × 1010 PFU Ad-hNGFβ 10 μl and 1.12 × 1010 PFU Ad-LacZ 10 μl respectively after pain threshold was measured on 21d after STZ injection.The mechanical and thermal pain threshold was measured before STZ injection (baseline) and at 21,35 and 49 d after STZ injection.The expression of SP in the dorsal root ganglion was determined after the measurement of pain threshold on 49 d after STZ injection.Results Compared with group C,the mechanical and thermal pain threshold was significantly decreased at each time point after STZ injection in groups DNP,Ad-NGF and Ad-LacZ,and the expression of SP in the dorsal root ganglion was signilicantly downregulated in groups DNP and Ad-LacZ (P ＜ 0.05).Compared with group DNP,the thermal pain threshold was significantly increased on 49 d afar STZ injection,and the expression of SP in the dorsal root ganglion was significantly up-regulated in group Ad-NGF ( P ＜ 0.01 ),and no significant change was found in the mechanical and thermal pain threshold and the expression of SP in the dorsal root ganglion at each time point in group Ad-LacZ ( P ＞ 0.05).Conclusion Subcutaneous transfection of Ad-hNGFβ can attenuate DNP to some extent through upregulation of the expression of SP in rat dorsal root ganglion.

Objective To investigate the effects of adenovirus containing human beta-nerve growth factor (Ad-hNGFβ) gene on substance P (SP) and calcitonin gene-related peptide (CGRP) content of the spinal cord in a rat model of neuropathic pain by chronic constrictive injury (CCI). Methods Forty-eight male SD rats weighing 200-250 g were randomly divided into 3 groups (n=16 each) : group Ⅰ sham operation; group Ⅱ CCI and group Ⅲ CCI + Ad-hNGFβ gene IT. The animals were anesthetized with intraperitoneal choral hydrate 300-350 mg/kg. The right sciatic nerve was exposed and 4 ligatures were placed on the sciatic nerve at 1-2 nun intervals as described by Bennet and Xie[5]. In sham operation group, right sciatic nerve was exposed but not ligated. In group Ⅰ and Ⅱ artificial cerebrnspinal fluid was injected IT instead of Ad-hNGFβ gene. The behavior score and the paw-withdrawal latency (PWL) to radiant heat and mechanical stimulus were measured one day before operation and every 4 days within the 28 days after gene transfection. Four animals were killed at 4, 7, 14 and 28 day after IT gene transfection in each group and lumbar segment (L4-6 ) of the spinal cord was removed for determination of SP and CGRP content by immunohistochemistry. Results The behavior scores were significantly higher and PWL to radiant heat and mechanical stimulus were significantly lower in group Ⅱ and Ⅲ than in group Ⅰ. There was no significant difference in the behavior score and PWL to mechanical stimulus between group Ⅱ and Ⅲ while the PWL to radiant heat was significantly higher in group Ⅲ than in group Ⅱ. After operation SP and CGRP content were significantly higher in group Ⅱ and group Ⅲ than in group Ⅰ , and significangly lower in group Ⅲ than in group Ⅱ 7-28 days after operation. Conclusion The recomhinant Ad-hNGFβ gene transfection can attenuate heat hyperalgesia by reducing SP and CGRP content of the spinal cord in a rat model of neuropathic pain induced by CCI.