Parkinson′s disease(PD)is a common disease caused by multiple factors and characterized by pathological degen?eration in the dopaminergic neural system. Based on its pathogenic factors,PD can be divided into several subtypes,so it is essential to develop therapeutic agents based on the main pathogenic factor of each subtype of PD. Recently it is confirmed that the mutation of leucine-rich repeat kinase 2(LRRK2)gene leads to increased activity of the LRRK2 notably,and then causes neurodegeneration. Thus developing LRRK2 inhibitors to modulate the kinase activity will be a novel therapy for the PD subtype which is caused by LRRK2 gene mutation. LRRK2,either a kinase or a GTPase,has two drug binding sites. Therefore,two types of LRRK2 inhibitors are being studied,one is the kinase inhibitor and the other is GTPase inhibitor. This paper summarizes the recent progress in the dis?covery and development of LRRK2 inhibitors.

Objective To understand the effects of NO signal system on the ciliary beating frequency (CBF) of airway epithelial cellMethods Nine normal male Sprague-Dawley rats were anesthetized with isoflurane Their tracheas were rapidly removed using aseptic technique The mucosa of trachea were cut into 1mm2 explants and cultured in DMEM The explants were divided into 5 groups as bellow: L-Arg group, 1-Hydroxy-2-oxo-3-(N-ethyl-2-aminoethyl)-3-ethyl-1-triazene (NOC-12) group, D-Arg group, 8-Br-cGMP group, and phosphate buffered saline (PBS)group Actively beating ciliated cells were observed, and their motion was quantified by measuring CBF using phase-contrast microscopy and videotape analysis Results L-Arg increased CBF from (7 43?0 75)Hz to(8 59?0 93)Hz (P

Objective To understand the modulating mechanisms in ciliary motility by NO signal pathway. Methods L arginine, the preferred substrate of NOS, was employed to act upon the cultured rat ciliated epithelia. After pre incubating with L NMMA, a NOS inhibitor, or ODQ,a sGC inhibitor, or Rp 8 Br cGMPS, an antagonist of PKG respectively, tissues were contacted with L arginine again. Ciliary beating frequency (CBF) was measured by phase contrast microscope and videotape analysis. Results L arginine increased CBF siginficantly. The effect of L arginine on CBF was blocked by L NMMA, ODQ or Rp 8 Br cGMPS.Conclusion L arginine may increase CBF via NO sGC cGMP PKG pathway.

Objectives:To investigate nutritional support in severely burned patients with delayed fluid resuscitation. Methods:From January 1990 through December 2000,62 cases with delayed fluid resuscitation were admitted to our burn department and were divided by different periods into two groups:group N(1990-1994,n=26) and group A(1995-2000,n=36).Group A was treated with recombinant human growth hormone(rhGH),early enteral feeding(EEF) and glutamine(Gln). Plasma albumin,pre-albumin,insulin,blood glucose and urine glucose levels were measured and lymphocyte was counted immediately after hospitalization and postburn day(PBD) 1,3,7,14,21,28. Results:①The survival rate in group A was very significantly higher than in group N.The complication in group A was significantly lower than in group N.②The time of wound healing in group A was shorter than in group N.③Plasma albumin,pre-albumin levels and lymphocyte count were decreased in two groups and was more serious in the group N(P

Objective To evaluate the efficacy of the combined saphenous nerve-great saphenous vein flap and cutaneous branches of posterior tibial artery flap in repairing refractory wounds. Methods Eighteen cases of pedal chronic ulcers were treated with the combinedsaphenous nervegreat Saphenous vein flap and cutaneous branches of posterior tibial artery flap, in which the wounds were treated with vacuum suction techniques before the operation in 6 cases. Wounds were from 8 cm× 13 cm to 1 cm× 17 cm in zine after debricement, and the designed size of the flaps was from 8 cm× 14 cm to 11 cm× 18 cm. Results After the treatment, 18 cases were evaluated as excellent in 10 cases, and good in 8 cases, in which the primary sealing of the wounds was achieved in 17 cases, but one case presented with focal necrosis of smaal size owing to vein drainage disturbance in a distallypedicled flap, and was healed after flap transplantation. Follow-up for 6 months to 2 years showed that all the patients were satisfied with the results. Conclusions The combined flap has reliable blood supply, skin pedicle of the flap is longer, superior texture and satisfied appearance, and incisive area of the flap is larger. It is particularly useful in repairing refractory wound in foot.

Objective To investigate the clinicopathological difference and prognosis of colorectal adenocarcinomas including mutinous, Signet-ring cell, papillary and tubular carcinomas. Methods Two thousand and eighty-nine patients with colorectal cancer underwent colorectal operation between August 1994 and April 2007. The clinicopathological characteristics of mucinous adenocarcinoma (n=144), signet-ring cell carcinoma (n=25), papillary and tubular carcinomas (n= 1837) were compared expect of other types of cancer (n = 83). The single factor and Logistic regression methods were used to analyze the clinicopathological parameters that influence the prognosis of colorectal cancer such as age, location of the tumor, staging, peritoneum and pathological typing. The survival rates of patients with above three types of adenocareinomas were analyzed. Results The mean age of onset was lowest in patients with mutinous adenocarcinomas [(54. 20 ± 16.25) years] compared with that in patients with signet-ring cell cancer [(40.43 ± 12.88)years] or papillary and tubular carcinomas [(58. 73 ±13.62)]. There were significant differences in gender, size and location of the tumor, TNM staging, peritoneal metastasis, lymph node involvement and adjacent organ invasion among three groups (all P values <0.05). The single factor and Logistic regression analysis revealed that both mucinous adenocarcinoma and signet-ring cell carcinoma were risk factors ot prognosis. The patients with mucinous adenocarcinoma or signet-ring cell tumor were poor in long-term overall survival in comparison with patients with papillary and tubular carcinoma (P<0. 001). Conclusions The colorectal mucinous and signet-ring cell adenocarcinomas are risk factors for prognosis of colorectal cancer, which imply the poor outcome.

Objective To investigate the efficacy of sunitinib in the treatment of patients with imatinibresistant advanced gastrointestinal stromal tumor (GIST).Methods The clinical data of 45 patients with imatinib-resistant advanced GIST who received the treatment of sunitinib (37.5 mg/d) at the First Affiliated Hospital of Sun Yat-Sen University from March 2008 to June 2012 were retrospectively analyzed.The mutation of c-kit and platelet-derived growth factor receptor α (PDGFRα) was detected,and the efficacy of imatinib was assessed after the treatment for 3 months,and factors influencing the survival were analysed.The survival rate was calculated using the Kaplan-Meier method,survival analysis was done using the one-way analysis of variance,and multivariate analysis was done using the COX regression model.Results The median time of treatment with sunitinib for the 45 patients was 11.0 months (range,4-37 months).The complete remission rate,partial response rate,rate of stabilized condition and disease progression rate were 15.6％ (7/45),8.9％ (4/45),46.7％ (21/45) and 28.9％ (13/45) after the treatment with sunitinib for 3 months.All the patients with clinical (imaging) complete remission received surgery for metastatic lesions or B-ultrasound guided ablation for single liver metastasis before the treatment with sunitinib.The most common grade 3 or 4 adverse reactions of sunitinib were hand-foot syndrome and anemia.C-kit and PDGFRα mutational analysis were carried out.C-kit exon 9 mutation was detected in 9 patients,c-kit exon 11 mutation in 21 patients,and no mutation was detected in 12 patients.The median progression-free survival time was 8.0 months (range,4.1-11.9 months),and the median overall survival time was 25.0 months (range,13.4-36.6 months).The results of univariate analysis showed that the primary lesion sites and mutational status of primary lesions were factors influencing the progression-free survival and overall survival (x2=5.967,6.622 ; 7.965,8.765,P ＜ 0.05).The results of multivariate analysis showed that only the mutational status of c-kit of primary lesions was the independent factor influencing the progression-free survival and overall survival (Wald =6.540,7.205,P ＜ 0.05).The progression-free survival and overall survival of patients with c-kit exon 9 mutation and patients with no gene mutation were significantly longer than patients with c-kit exon 11 mutation (x2 =7.965,8.765,P ＜ 0.05).Conclusion Sunitinib with a dosage of 37.5 mg/d could effectively treat patients with imatinib-resistant advanced GIST.A better survival is observed in patients with c-kit exon 9 mutation or with no gene mutation when compared with patients with c-kit exon 11 mutation.

Objective To determine the effects of histone deacetylase inhibitor suberoylanilide hydroxamic acid(SAHA) on the cell proliferation and apoptosis of the human hepatic stellate cell line LX-2.The possible underlying mechanisms were also investigated.Methods The LX-2 cells were treated with SAHA in vitro.The morphology of LX-2 cells in different concentrations groups was observed by inverted microscope;the proliferation of LX-2 cells was measured by MTT assay;the Annexin V-FITC and PI staining was used to detect the apoptosis of LX-2 cells by flow cytometry and fluorescence microscope;the expression of α-SMA,collagen Ⅰ,acH3K9,acH3K14 and acH3K18 were detected by Western blot.Results The morphology change of LX-2 cells showed that SAHA inhibited the proliferation rate of LX-2 cells and in a dose dependent manner(P<0.05).The LX-2 cells were sensitive to SAHA along with time increasing,and in a time-dependent manner(P<0.05).Western blot showed that the expression levels of α-SMA and collagen-Ⅰ were significantly lower(P<0.05),on the contrary,the acetylation levels of acH3K9,acH3K14 and acH3K18 were significantly higher (P<0.05).Conclusions The increased acetylation of the histone acH3K9,acH3K14,acH3K18 and the lower expressed α-SMA and collagen-Ⅰ in LX-2 cells may be one of the mechanisms of SAHA.

ObjectiveTo analyze the clinicopathological features and prognosis of gastric cancer patients with metastatic nodules of perigastric soft tissue. MethodsIn this study,1025 cases of gastric cancer received radical resection.According to the metastasis of perigastric soft tissue,patients were divided into metastatic group ( group MP,n =334 ),non-metastatic group ( group NMP,n =691 ).The clinicopathological features and prognosis were compared between the two groups. ResultsIn group MP,the ratio of upper,middle,lower,total gastric cancer was 25.8％,22.0％,51.4％,0.9％ and the ratio in group NMP was 33.2％,21.3％,41.3％,4.2％ respectively,showing significant higher ratio of upper and total gastric cancer in MP group(P =0.000). In group MP 47.3％ cases with tumor size ≥5 cm,significantly higher than that in NMP group(27％ ) (P =0.000).Lymph node metastatic ratio between 21％ -40％ and 41％ -100％ was found in 24.4％ and 37.3％ in MP group respectively,significantly higher than that of 12.9％,10.8％ in NMP group(P =0.000).20.1％ cases had distal metastasis in group MP,significantly higher than that of 4.1％ in group NMP(P=0.000).In group MP and NMP group,the ratio of Borrmann infiltration typing was 82.1％ vs.64.6％,the ratio of positive CEA was 21.2％ vs.11.4％,the ratio of lower or undifferentiation typing was 78.7％ vs.64.2％,all with significant difference (P =0.000 ). COX regression analysis showed the infiltration depth,organic invasion,lymph node metastatic ratio,M staging,Borrmann typing,metastatic nodules was the independent prognostic factors.Prognosis was significantly poorer in the cases with perigastric soft tissues than without ( P =0.000 ).Stratified analysis showed that irrespective of tumor size,infiltration depth,lymph node metastatic ratio,CEA value,Borrmann typing,differentiation degree,the mean survival time was significantly shorter in MP group than that in group NMP(P ＜ 0.005).In cases without distal metastasis,the prognosis was significant poorer in group MP than that in group NMP ( P =0.000 ),however,there was no significant difference between two groups in cases without distal metastasis ( P =0.076).ConclusionsPerigastric soft tissue metastasis was common in gastric cancer,more frequently seen in tumor ≥5 cm,or with organic invasion,lymph nodemetastaticration ≥ 21％, distalmetastasis, Borrmanninfiltrationtyping, loweror undifferentiation typing,positive CEA. Perigastric soft tissues metastasis was the independent prognotic factor for gastric cancer.

AIM:To study the effects of suberoylanilide hydroxamic acid (SAHA) on the apoptosis of hepatic stellate cells (HSCs) and expression of associated proteins, and to investigate the mechanisms of SAHA to induce apoptosis.METHODS:The rat HSCs were isolated by OptiPrep gradient centrifugation method.The effect of SAHA on HSC proliferation was detected by real-time cell analyzer.The morphological changes of HSCs treated with SAHA at different concentrations were observed under inverted microscope.The apoptotic rates of HSCs were analyzed by flow cytometry with Annexin V-FITC/PI staining and fluorescence microscopy.The protein expression of α-smooth muscle actin (α-SMA), collagen I, tissue inhibitor of metalloproteinase 1 (TIMP1), glucose-regulated protein 78 (GRP78) and histone deacetylase 6 (HDAC6) was detected by Western blotting.The interaction of GRP78 with HDAC6 in the HSCs was determined by co-immunoprecipitation.RESULTS:HSCs were successfully isolated and cultured for 14 d, during which the HSCs changed gradually from rest state to active state.SAHA significantly inhibited the proliferation of HSCs in a time-and dose-dependent manner (P<0.05).The results of Western blotting showed that the protein expression levels of α-SMA, TIMP1, collagen-I and HDAC6 were significantly decreased (P<0.05), while GRP78 was significantly increased (P<0.05).Compared with activated HSCs, GRP78 and total acetyl-lysine protein were significantly increased in the co-immunoprecipitated HSCs treated with SAHA, while HDAC6 protein was significantly decreased, indicting that GRP78 formed a complex with HDAC6.CONCLUSION:The anti-hepatic fibrosis effect of SAHA may be related to down-regulation of HDAC6 and up-regulation of acetylated GRP78, thus inducing endoplasmic reticulum stress of HSCs and promoting the apoptosis of HSCs.