Objective To discuss the clinical effect of behavioral therapy combined with pramipexole in patients’ degree of depression and non-motor symptoms in early-onset Parkinson’s disease and depression.Methods A total of 74 cases of patients with early-onset Parkinson’s disease and depression were equally divided into observation group and control group, 37 cases in each groups.Patients in control group were given pramipexole, while patients in observation group were given pramipexole and behavioral therapy.The hamilton depression ( HAMD) scale, Zung self-rating depression scale and unified Parkinson’s disease rating scale ( UPDRS ) were used to evaluate and measure the change of degree of depression and non-motor symptoms.Results Before treatment, the HAMD score, Zung score, UPDRS II score and UPDRS III score between two groups had no statistical difference; after treatment, the HAMD score, Zung score, UPDRS II score and UPDRS III score in two groups were significantly decreased (P<0.05). The HAMD score, and Zung score between two groups had no statistical difference at the end of 4th weekends, and compared with control group, those scores in observation group were much better at the end of 8th and 12th weekend (P<0.05).The UPDRS II and UPDRS III between two group had no statistical difference at the end of 4-8th week, while those scores in observation group were better than those in control group at the end of 12th week(P<0.05).Conclusion Behavioral therapy combined with pramipexole has a great effect on the improvement of patients’ degree of depression and non-motor symptoms, which has a positive promotion on patients’ life quality.

Objective To assess the safety and the efficacy of endoscopic ultrasound (EUS)-guided transmural drainage of pancreatic pseudocysts (PPC).Methods A total of 17 patients with PPC who underwent EUS to detect the optimal site and depth of puncture.The needle was punctured into the PPC cavity through endoscopic biopsy hole,cyst fluid was drained with a syringe.The guide wire was inserted along the pinhole under X-ray,and then the needle-knife was sent along the guide wire to cut the gastric wall and pseudocysts wall,followed by balloon dilation.The way of drainage was selected according to the cyst fluid properties.The technical success rate,treatment success rate,complication occurring rate and the skills were evaluated.Results Four patients were with nasalcystic catheter drainage,9 patients with double pigtail stents internal drainage,and 4 patients with nasal-cystic catheter and double pigtail stents combination drainage.The treatment success rates were 3/4,7/9,and 4/4 respectively.Only 1 patient subsequently developed bleeding from puncture site after stent successively placed,and was turned to surgery because of ineffective endoscopic treatment.Infection occurred in 4 patients during drainage,two of those were switched to surgical resection due to poor medical treatment response,and the other 2 were cured with intravenous infusion of antibiotics sensitive to cyst fluid bacteria and metronidazole rinse PPC.The median follow-up duration was 28.5months,and there was none of recurrence.Conclusions EUS-guided transmural drainage of PPC is safe.Stent placement and nasal-cystic catheter play an important role in PPC treatment.

BACKGROUND:Basic research demonstrated that type Ⅰ collagen exhibited prominent effect on osteogenesis,bone mass and bone fracture,which also participated in the bone fusion.However,few reports concerning the polymorphism of type Ⅰ collagen gene and spinal fusion.OBJECTIVE:To investigate the polymorphism of type Ⅰ collagen and to explore its relationship with the spinal fusion rate following metal implant or autogenous bone transplantation.METHODS:A total of 200 volunteers who need to receive spinal fusion in the Affiliated Hospital of Qingdao University Medical College were selected,including 102 cases received anterior cervical subcorpectomy combined with lilac bone implantation fusion following decompression,and 98 cases received posterior laminectomy for decompression combined with intertransverse process fusion.Meantime,223 normal adults were served as the control group.The peripheral blood was drawn-off and genomic DNA was extracted from white blood cells.The specific fragment which includes the objective gene was amplified by polymerase chain reaction (PCR),with length of 293 bp.The genotypes of Pcol2 site in type Ⅰ collagen were detected by PCR-restriction fragment length polymorphism (PCR-RFLP) method.The PCR product was digested with restriction endonuclease Eco311 and the result was observed by agarose gel electrophoresis.The G gene represented for the presence of the restriction endonuclease site,while the T gene for the absence of the restriction endonuclease site.The fusion rate of the bone graft was evaluated by x-ray film prior to and at months 3,6 and 12 after operation,and the results were compared by stages including quick (＜3 months),middle (3-6 months) and slow (6-12 months).RESULT AND CONCLUSION:There were the-1997G/T polymorphisms of the type Ⅰ collagen gene in 423 cases,including 166cases with GG,232 cases with GT,and 25 cases with TT,in addition,there was some correlation between the GG genotype and the lilac bone implantation fusion (P =0.004).The GG genotype accounted for 50% in the fast group,which was obviously greater than that of the middle and slow groups (33.3% and 16.7%,respectively).However,the-1997G/T polymorphisms had no correlation with the bone graft fusions inter transverse process of lumbar vertebra (P=0.831).The GG genotype in the-1997G/T polymophsim of the type Ⅰ collagen gene may be the essential factor which can promote the C-spine auto-ilium graft fusion.

Objective To investigate the effect of active vitamin D (VD) on macrophage M1 and M2 phenotype and its role in protecting podocyte impairment in diabetic nephropathy (DN).Methods Diabetes mellitus rats were established by intraperitoneal injection with streptozocin.Rats were randomly divided into four groups:normal-1 (NC-1,n=8),normal-2 (NC-2,n=8,normal rats treated with calcitriol 0.1 μg· kg-1 · d-1 by gavages),DN (n=24) and VD (n=24,DN+calcitriol 0.1 μg· kg-1 · d-1 by gavages).Blood glucose and body weight were assessed,and 24-hour urine was collected regularly.Blood and urine samples were taken for biochemical study,and kidney tissues were used for PAS staining to assess histological changes.Immunohistochemical staining was used to detect number of CD68 + macrophage.Western blotting was used to detect protein expressions of nephrin,podocin,CD68,M1 specific marker of inducible nitric oxide synthase (iNOS),TNF-α and M2 specific marker of CD163,arginase 1 (Arg-1),mannose receptor (MR).Results (1) In DN group,levels of BUN,Scr,urinary protein and glomerular mesangial matrix proliferation were significantly higher (P ＜ 0.05),and the expressions of nephrin,podocin were significantly decreased compared with NC groups (P ＜ 0.05).These above changes were significantly improved in VD group (P ＜ 0.05).(2)Number of CD68 + macrophage infiltration in DN group was increased in a time dependent manner compared with NC groups,which was significantly reduced in VD group (P ＜ 0.05).(3)To further definite M1 and M2 macrophage activation phenotype,the protein expressions of iNOS and TNF-α was increased in DN group at 8th,14th,18th weeks compared with NC groups (P ＜ 0.05),which were significantly decreased in VD group (P ＜ 0.05).Although,there were no significant difference of protein expressions of CD163,Arg-1 and MR between VD and DN group at both 8th and 14th week (P ＞ 0.05),the protein expressions of CD163,Arg-1 and MR were higher in VD group at 18th week than that in DN group (P ＜ 0.05),and the ratio of CD163/CD68 was also enhanced in VD group (P ＜0.05).(4)Moreover,the protein expression of iNOS was negatively correlated with expression of either nephrin or podocin (r =-0.707,P ＜ 0.01; r =-0.712,P ＜ 0.01),whereas the protein expression of CD163 was positively correlated with expression of either nephrin or podocin (r =0.627,P＜ 0.01; r=0.613,P ＜ 0.01).Conclusion Vitamin D can regulate macrophage phenotype,via inhibiting M 1 macrophage activation and enhancing M2 macrophage activation to protect podocyte impairment.

Objective To investigate the relationship between a functional single nucleotide polymorphisms(SNP) in the core promoter region of GDF5 gene(+104T＞c ;rs143383) and spinal fusion in Qingdao Han people. Methods This study included 201 patients who needed to be treated with spinal fusion and 200 healthy controls. They were all out of tuberculosis, tumor, infection, and long time of related medicine using. Their operation regions were similar, containing L4.5, C4.5, C5.6, and C6.7. The SNP was defined in all people, using PCR-restriction fragment length polymorphism (RFLP) and gene sequencing. The patients were followed-up 3,6, 12 months after operation. The conditions of bone graft fusion were carried out into different grades according to imaging. According to the common used fusion criterion, to analyze the relationship between this SNP and spinal fusion. Results The results obtained from PCR-RFLP were confirmed by gene sequencing. The patients and the healthy control all showed the SNP in this site. There were no relationship between the spinal fusion patients and the healthy control in the SNP (x2=0.304, P=0.859). But it showed correlation (x2=4.752, P=0.023) with fusing or not in the spine and the speed of fusion (x2=9.864, P=0.007)in Qingdao Han people. In the fusion group, the site rs143383 showed more C allele than the non-fusion group. T allele may affect the transcription of GDF5, thus reduce the expression of GDF5. Patients with the genotype TC+CC showed larger proportion stable fusion and faster speed than the patients with TT. Conclusion SNP in the core promoter region of GDF5 (+104T＞C) is associated with spinal fusion in Qingdao Han people. The allele C may be an important factor to promote spinal fusion. Detect the TT genotype early and intervene, the spinal fusion effect may be improved. Or, the genotype may be changed by gene technology,making the efficient fusion.

Objective To investigate the cytotoxicity and radiosensitization of 5-nitroindazole-3-formyliminodiacetic acid on HeLa cells.Methods HeLa cells in exponential growth phase were incubated in culture media with different doses and the survival rate was determined by MTT assay.The survival rate of cells receiving radiation combined with different doses of medicine was compared with that of the control.Results The cytotoxicity of S-nitroindazole-3-formyliminodiacetic acid on HeLa cells was very low.The drug had hypoxia radiosensitizing effect on HeLa cells.At doses of 0,6,12,24,48 and 96 μg/ml under hypoxia ,the survival rate were 0.91 ,0.87,0.84,0.81 ,0.76 and 0.60,respectively.At the dosage of 48 and 96 μg/ml,the survival rate were 0.85 and 0.73 under oxygenous).Conclusions 5-Nitroindazole-3-formyliminodiacetic acid has low cytotoxicity and rediosensitizing effect on HeLa cells.

Objective To analysis curative effect of triple therapy and sequential therapy for eradication of Helicobacter pylori in patients with long-term aspirin use.Methods 52 patients with long-term aspirin use who were diagnosed with helicobacter pylori infection were collected.All patients were divided into sequential group and triple group according to different drugs classified, each 26 cases in each group were given corresponding drug treatment, after the end of treatment, the serum pepsinogen, Hp-IgG antibody levels and Helicobacter pylori clearance rate were detected in all patients. Results After treatment, compared with the triple Group, in the sequential group, the levels of serum PG I and PG II were lower, and PGⅠ/PGⅡwas higher, and the differences were statistically significant (P<0.05);the serum Hp-IgG antibody levels were lower in the sequential group (P<0.05);the Hp clearance rate of patients in the sequential group was higher (P<0.05).Conclusion Compared with triple therapy, sequential therapy can lower serum PG, PG II and Hp-IgG levels, and improve the Hp clearance rate and clinical efficacy, have guiding significance to clinical.

Objective To investigate the effect of 1,25(OH)2D3 on high glucose induced podocyte injury and its signal transduction mechanism.Methods Differentiated mouse podocytes were exposed to normal glucose,high glucose,and different concentrations of 1,25(OH)2D3 or LY294002 (a selective PI3K inhibitor) for 24 h.PCR and immunofluorescent staining were used to detect nephrin,podocin,and desmin.Western blotting was used to detect protein expression of nephrin,podocin,desmin,PI3K,Akt and p-Akt.Results Compared with high glucose group,1,25(OH)2D3 (100 nmol/L and 1000 nmol/L) significantly up-regulated the expression of podocin and nephrin in podocytes induced by high glucose (P ＜ 0.05).Meanwhile,1,25(OH)2D3 (100 nmol/L) significantly reduced the expression of desmin (P ＜ 0.05).PI3K and p-Akt were obviously reduced in high glucose group.In the presence of 1,25(OH)2D3,the trends were reversed.However the above effects of 1,25(OH)2D3 were abolished when p-Akt was blocked by the PI3K inhibitor LY294002.Conclusions 1,25 (OH)2D3 can inhibit high glucose-induced pedocyte injury through PI3K/p-Akt signaling pathway.

Objective To investigate the effects and underlying mechanism of calcitriol on ameliorating podocytes impairment in DN rats.Methods SD rats were randomly divided into four groups:normal control (NC) group,calcitriol treatment (VD) group:calcitriol 0.1μg· kg--1 d-1,diabetic nephropathy (DN) group:streptozocin (STZ) 58 mg/kg,DN treated with calcitriol (DN + VD) group:calcitriol 0.1 μg · kg-1 · d-1 + STZ 58 mg/kg.Rats were sacrificed at the end of 18 weeks.Results Compared with the DN group,the DN + VD group exhibited significantly lower proteinuria by 36％,improved renal histology at the end of the experiment (P ＜ 0.05),and similar levels of blood glucose,serum urea nitrogen as well as body weight (P ＞ 0.05).There were no significant differences in the serum concentrations of creatinine,calcium and phosphorus among the four groups (P ＞ 0.05).In DN group,the expressions of nephrin,podocin,VDR,PI3K-p85 and p-Akt were significantly decreased and the expression of desmin was increased compared to NC group.Calcitriol treatment could attenuate the above changes.Additionally,a positive correlation was observed between the expressions of nephrin and VDR (r=0.776,P ＜ 0.05).Likewise,the expression of nephrin was positively correlated with either PI3K -p85 or p-Akt (r=-0.736,r=0.855,all P ＜ 0.05).Conclusion Calcitriol can ameliorate podocytes injury in DN rats,which might be related with the further up-regulation of PI3K/p-Akt signaling pathway.

Objective To investigate the effect of 1,25(OH)2D3 on high glucose induced macrophage activation and its underlying signal transduction mechanism.Methods RAW 264.7 cells were used to perform cell culture,the activity of intracellular iNOS was measured.VDR siRNA and PPARγ antagonist pre-treatment with macrophages were done before using 10-8 mol/L1,25(OH)2D3 to intervene high glucose pre-incubated macrophages.M1 markers including iNOS,TNF-α,IL-12,M2 markers including MR,Arg-1,IL-10 and nuclear receptors VDR and PPARγ were separately examined.Results The iNOS activity was increased in a glucose-dose and time dependent manner.Particularly,25 mmol/L glucose at 24 h gave the maximum response.After being treated with 25 mmol/L glucose for 24 h,not only inflammatory cytokines of TNF-α,IL-12 in the supernatant were increased,but quantitative real-time PCR and Western blotting analysis showed iNOS was also up-regulated (P ＜ 0.05).However,M2 markers,i.e.MR and Arg-l were significantly decreased (P ＜ 0.05).When in the presence of 1,25(OH),D3,the trends were reversed:the markers of M1,including TNF-α,IL-12 and iNOS were obviously reduced (P ＜ 0.05),while M2 markers,IL-10,Arg-1 and MR were increased (P ＜ 0.05).In addition,VDR and PPARγ were also increased (P ＜ 0.05).However,the above effects of 1,25 (OH)2D3 were abolished when further inhibited the expression of VDR and PPARγby VDR siRNA and PPARγ antagonist.Besides,accompanied by VDR,PPARγwas also decreased upon the treatment with VDR siRNA (P ＜ 0.05).Conclusion 1,25(OH)2D3 can promote high glucose induced classically activated macrophages (M1) converting to alternatively activated macrophages (M2) and this is achieved through VDR-PPARγ pathway.