Parkinson's disease(PD) is a common neurodegenerative disorder with no effective protective treatment,characterized by a massive degeneration of dopaminergic neurons in the substantia nigra(SNpc) and the subsequent loss of their projecting nerve fibers in the striatum.The major neurochemical manifestation of this disorder is the loss of the neurotransmitter dopamine(DA) in the striatum as a result of the progressive degeneration of the dopaminergic neurons in the substantia nigra.There have been significant progresses in recent years reporting on the use of mesenchymal stem cells(MSCs)in gene therapy,with specific application towards PD.MSCs,a kind of multipotent adult progenitor cells,are considered as a useful vehicle for cell and gene therapy because of their multiple differentiation potentiality and self-transplantation.The present study was focused on treating rat model of PD using human tyrosine hydroxylase gene(hTH),human aromatic L-amino acid decarboxylase gene(hAADC) and human GT Pcyclohydrolase I gene(hGCH-I) engineered MSCs,in order to provide a better understanding about the application of these cells in the therapeutic benifit of PD.The gene of hTH,hAADC and hGCH-I were introduced via recombinant adeno-associated virus(rAAV) infection into the MSCs in vitro.The genetically modified MSCs expressing hTH,hAADC and hGCH-I were transplanted into the striatum of PD rat models.The behavior,the nigra-striatal level of DA and its metabolite were detected.The results of present study were shown as follows:hTH,hAADC,hGCH-I and LacZ gene were transfected into MSCs with adeno-associated virus vectors.The HEK293 packaging cells(ATCC) were transfected with the plasmids of pAAV-hTH,pAAV-hAADC,pAAV-hGCH-I,pAAV-LacZ,pAAV-RC,pHelper by using calcium phosphate precipitation.Titer was detected using HT1080 cells.Viral particles were collected and used to infect MSCs.The purified modified MSCs expressing the three kinds of genes were selected separately and were grafted in the striatum of the PD model rats in the lesion side.The MSCs genetically modified suvived well 12 weeks after transplantation.The improvements of the behavior were observed every week after transplantation.Compared with the control group,the rounds of asymmetric rotation after apomorphine administration decreased in the groups double or triple genes engineered MSCs grafted(p

Objective To evaluate the feasibility and safety of double-lumen balloon catheter applied in patients with achalasia of cricopharyngeal muscle. Method Fifty patients with achalasia of cricopharyngeal muscle were randomly divided into experimental group and control group. All the patients received routine drug treatment,swallowing function training,feeding training and low frequency VitalStim electric stimulation. In addition,double-lumen balloon catheter and #14 urinary catheters were applied to patients in the experimental group and control group,respectively. The swallow water tests and video fluoroscopy swallowing study(VFSS) were used to evaluate the treatment effects,the electron-nasopharyngolaryngoscope was used to assess bleeding and swelling of mucous membrane,and VRS-5 was used to assess pain. Result After treatment,the scores of swallow water tests and VFSS were significantly better than those before treatment in both groups(P<0.05). There was no significant difference between the two groups(P>0.05). However,the incidence of complications was significantly higher in the control group than that of experimental group(P<0.05). Conclusion Both treatment methods can effectively relieve the achalasia of cricopharyngeal muscle,but modified double-lumen balloon catheter can reduce the incidence of complications.

Objective To observe the effect of nutritional status change on hemodialysis patients of chronic renal failure and deficiency of spleen and kidney treated bytraditional Chinese medicine(TCM) of warming kidney and invigorating spleen. Methods In our center, 146 patients who were diagnosed chronic renal failure belonging to deficiency of spleen and kidney in TCM were received hemodialysis between Jun2012 and May2013. All patients were randomly divided into a treatment group (n=72) and a control group (n=74). 74 patients in the control groupreceived conventional hemodialysis and medicine treatment, and 72 patients in treatment group received conventional hemodialysis、medicine treatment and TCM treatment of warming kidney and invigorating spleen. The comprehensive nutritional assessment of all patients were treated by modified SGAN method (MQSGAN), anthropometric, biochemical and laboratory examination before and 6 monthsafter the treatment. Results The treatment group patients of modified SGAN(9.58±3.15), BMI (body mass index 19.34±0.52) Kg/m2, MAC(upper arm circumference 27.51±1.95)cm, TSF(tricepsskinfold thickness 12.92± 2.42) mm and MAMC(upper arm muscle circumference 23.64±1.96)cm were significantly improved compared with the control g roup(respectively data 13.23±3.14, 17.29±0.76) Kg/m2, (24.01±2.55, 10.58±2.71) mm, (20.71±2.04)cm, P<0.05,and treatment group patients with ALB(serum albumin 38.19±1.95)g/L, PA(prealbumin 2.23±0.16)g/L, TF(transferrin 0.21±0.04)g/L, CH(cholesterol 4.02±0.26)mmol/L, BUN(blood urea nitrogen 19.58±2.17)mmol/L and SCr (creatinine 869.54±79.15)mmol/L were significant improved also compared with the control group(respectively data 33.73±1.31)g/L, (1.67±0.25)g/L, (0.17±0.02)g/L, (3.22± 0.46)mmol/L, (16.27±1.12)mmol/L, (792.73±71.65)mmol/L, P<0.05. Conclusion Traditional Chinese medicine of warming kidney and invigorating spleen can improve the nutritional status and quality of life on patients with chronic renal failure belonging to deficiency of spleen and kidney type during hemodialysis.

AIM: To investigate the relation between the changes of the cerebral microvasculature and the expression of plasminogen activator inhibitor-1 (PAI-1) in ischemic focal and perifocal areas after 2 hours focal cerebral ischemia with reperfusion in rats. METHODS: The changes of cerebral microvascular structure were observed by optical microscope and electric microscope, the expression of PAI-1 were assessed by immunohistochemical staining, Western blotting in ischemic focus and perifocal areas after focal cerebral ischemia with reperfusion. RESULTS: The edema of extra-cellular matrix and the hemorrhage of extravessels in ischemic focus and perifocal areas were most severe, and degradation and the defect of basement membrane were also observed after 6 hours and 3 days reperfusion following focal cerebral ischemia. The expression of PAI-1 decreased significantly compared with control group (P

ObjectiveTo observe the expressions of PPAR-γ protein during the course of pancreatic fibrosis of chronic pancreatitis (CP) in Wistar rats and its significance.Methods Bibutyltin dichloride ( DBTC,8 mg/kg body weight) in a volume of 200 ml solvent was injected into the tail vein to establish the CP rat's model.Wistar rats were randomly divided into control group and 1,3,5,7,14,42 d group according to weights.Pancreatic tissue underwent routine pathological examination,and collagen accumulation in pancreatic sections was determined by staining for Sirius Red.Pancreatic myeloperoxidase (MPO)activity was determined.Expressions of α-SMA and PPAR-γ proteins were assessed by immunohistochemical method.Results Light microscopy showed signs of acute pancreatitis with interstitial edema and infiltration of neutrophilic granulocytes 7 d after DBTC injection.Acinar cells necrosis,atrophy,lymphocytes and monocytes infiltration,fibrosis within lobule and peri-lobule as well as pancreatic duct changes were found,which was in accord with the course from AP to CP.One days after induction,the activity of MPO,expressions of α-SMA was significantly increased[ (0.78 ±0.71) vs (0.15 ±0.05)U/g,6.67 ±3.14 vs 0,P＜0.05],then it did not increase with time of induction.Seven days after induction,collagen level reached the peak [ (45.42 ±15.99)％ ],which was significantly higher than that in control group [ (10.87 ± 2.28 )％,P ＜ 0.05 ].Collagen fibers accumulated from periductal to intra-acinar and/or inter-acinar areas.In control rats,the expression of PPAR-γ protein was positive only in vessel walls,and the expression level was 0.17±0.41 and increased with time of induction,then reached the peak of 4.83 ± 2.71 at 42 d.ConclusionsDuring the course of pancreatic fibrosis in rats,the expression of PPAR-γ protein is gradually increased,and plays a limited anti-inflammation and anti-fibrosis role.

Aim To investigate the proliferation of HSCs stimulated by exogenous TGF-β1 (transforming growth factor betal),observe the effect of TFB(total flavonoids of Bidens Bipinnata L.)on smad2/7,typeⅠcollagen mRNA and protein expression of HSCs and study the protective effect and molecular mechanism of TFB on hepatic fibrosis.Methods HSCs were isolated with collagenase Ⅳ perfusion in situ and density gradient centrifugation. The effect of TFB on cell proliferation was observed by MTT colormetric assay. The auto-secretion of TGF-β1 and synthesis of type Ⅰ collagen were measured by enzyme-linked immuneadsordent assay (ELISA).Moreover,the expression of smad2/7, typeⅠcollagen mRNA and protein was measured by semi-quantitative RT-PCR and Western blot methods respectively.Results TFB could markedly inhibit the proliferation of HSCs of liver fibrosis rats stimulated by TGF-β1 and production of TGF-β1 and type Ⅰ collagen.In addition,TFB treatment could significantly down-regulate smad2 and type Ⅰ collagen mRNA expression and up-regulated smad7 mRNA expression of HSCs Smad2 protein expression of HSCs stimulated by TGF-β1 was also down-regulated by TFB.Conclusion TFB has the protective effect against hepatic fibrosis by inhibiting the activation of TGF-β1 signaling pathway and suppressing the HSC proliferation.

Objective To explore the diagnostic value of human telomerase reverse transcriptase (hTERT) and proliferating cell nuclear antigen (PCNA) in pleural and peritoneal effusion. Methods The expressions of hTERT and PCNA were detected by immunocytochemistry in 46 malignant and 21 benign serous specimens. Results hTERT and PCNA were detected respectively in 39 and 29 of 46 malignant specimens, but no expressions in reactive mesothelial cells in all serous specimens. hTERT correlated with PCNA (r=0.30, P

Objective To investigate the feasibility and safety of modified percutaneous left atrial appendage occlusion (PLAAO) under transthoracic echocardiographic (TTE) guidance without general anesthesia instead of transesophageal echocardiographic guidance.Methods A total of 14 patients who met the inclusion criteria underwent modified PLAAO guided by TTE instead of TEE without general anesthesia.Regular clinical follow-up observations of PLAAO-related major adverse events were done in the perioperative period.Results All patients were successfully implanted with left atrial appendage occluder device (Watchman) without device-related serious complications.Immediately occlusion success rate was 100％.No major adverse events occurred during hospitalization and follow-up.The mean operation time was 108 ± 22 min(range 75-150 min)and the mean radiation exposure time was 15.8 ± 7.6 min(range 8-32 min).Conclusion Modified PLAAO guided by TTE instead of TEE without general anesthesia may be safe and effective.This method simplifies the operation process and is favorable for PLAAO application.But this modified PLAAO is still needed to be validated in more patients.

Objective:To analyze the clinical outcome and feasibility for patients who underwent total parathyroidectomy without autotransplantation (TPTX) for secondary hyperparathyroidism (SHPT).Methods:From April 2012 to December 2015,220 SHPT patients underwent TPTX in the department of Breast and Thyroid Surgery of China-Japan Friendship Hospital.The clinical data and effect were assessed retrospectively.Results:All the 220 patients were on permanent dialysis with mean duration of dialysis (7.93 ± 3.75) years.A durable reduction in mean PTH,Ca and P were observed after TPTX (P＜0.01).The mean hospital stay was (7.8 ± 2.8) days.TPTX produced a rapid improvement in clinical symptoms.Incidence of hypocalcemia was 73.46％.Severe complications such as recurrent laryngeal nerve palsy or inactive dynamic osteopathia,haven't been observed postoperatively.The rate of persistent status (PTH≥300 pg/mL) was 9.1％.One (0.45％) died of infectious shock perioperatively.Conclusions:TPTX was a safe and feasible surgical procedure for patients with SHPT.It was worth of being applied.Not missing the parathyroid during operation was the key point for successful TPTX.Intensive monitoring and maintaining stable normocalcemia were the key point to reduce complication.

Background Researches demonstrated that dendritic cells(DCs) are uniformly immature in the central cornea but mature in the peripheral region of cornea.So an important question is which factor impact the maturation of DCs,especially in terms of corneal transplant rejection and the known roles of DCs in the development and persistence of some corneal diseases.Objective This study aimed to examine whether corneal stroma cells (CSCs) inhibit DCs maturation through secreting transforming growth factor beta 2 (TGF-β2) and prostaglandin E2 (PGE2).Methods DCs,T cells and CSCs were isolated and cultured from clean BALB/c and C57BL/6 mice.The level of PGE2 and TGF-β2in CSCs culture supernatant and the fresh RPMI 1640 medium were then analyzed by enzyme linked immunosorbent assay (ELISA).During the DCs maturation stage,the neutralizing TGF-β2 antibody and the EP2 receptor antagonist AH6809 were added in the CSCs culture supernatant respectively.According to the different treatment,cultured cells were assigned to different groups as follows:control group,CSCs culture supernatant group,AH6809 group,TGF-β2 antibody group,AH6809 +TGF-β2 antibody group.Subsequently,the cellular surface markers for DCs,including CD11c,CD80,CD86,and MHC- Ⅱ,were analyzed by flow cytometry.The capability of stimulating the proliferation of T lymphocytes was evaluated by allogeneic mixed lymphocyte reactions,and the function of endocytosis was assessed by fluorescein isothiocyanate-dextran(FITC) uptake.Results The data of ELISA showed a higher concentration of TGF-β2 and PGE2 in murine CSCs culture supernatant than in the fresh RPMI 1640 medium.Compared with the CSCs culture supernatant group,the expression of CD80,CD86,and MHC- Ⅱ was up-regulated ( P ＜ 0.05 ),the expression of dextran was down-regulated ( P ＜ 0.05 ),and the stimulate index was increased( P＜ 0.05 ) in the TGF-β2 antibody group; the expression of CD86,and MHC-Ⅱ was up-regulated (P＜0.05),the expression of dextran was down-regulated ( F =13.740,P =0.006 ),and the stimulate index was increased(P＜0.05) in the AH6809 group;the expression of MHC-Ⅱ was up-regulated and the stimulate index was increased with statistical difference in interaction(P＜0.05 ) in the AH6809+TGF-β2 antibody group.Compared with the control group,the expression of CD80 and CD86,and the stimulate index was still lower(P＜0.05 ).Conclusions TGF-β2 and PGE2 contribute to the inhibitory effects on DCs maturation mediated by murine CSCs in vitro and further have additive effect on the immunosuppression of DCs.