Objective To investigate the protein markers that specifically expressed in patients with acute rejection (ACR) after liver transplantation, and to explore preliminarily the mechanisms. Methods Serum samples from three patients with pathologically confirmed ACR after liver transplantation in Tianjin First Central Hospital were collected as ACR group. Three serum samples from patients with normal liver function indicators after liver transplantation were collected as No-ACR group. And six serum samples from healthy examination were mixed with equal amount as healthy control group. Isobaric Tags for Relative and Absolute Quantitation (iTRAQ) was employed to separate, screen and identify the differentially expressed proteins between three groups. KEGG and STRING software were applied to deeply analyze the data of three groups. Results A total of 88 differentially expressed proteins were found between ACR group and healthy control group. There were 39 differentially expressed proteins between No-ACR group and healthy control group. Ten differentially expressed proteins were acquired between ACR group and No-ACR group. Comparing 88 and 10 differentially expressed proteins, 9 proteins were the same. Among 88 differentially expressed proteins, 30 of them showed a direct interaction, and can be positioned in 13 signaling pathways based on KEGG and STRING software. Fourteen (46.67%) of the 30 proteins were located in the complement and coagulation cascade pathway. Among 39 differentially expressed proteins, which were detected between No-ACR group and control group, 10 proteins showed a direct interaction including 9 proteins concentrated in the complement and coagulation cascade pathway. Conclusion By proteomic analysis, nine differentially expressed proteins are obtained, which may be regarded as the candidate bio-markers for ACR early diagnosis after liver transplantation. The complement and coagulation cascades system is significantly adjusted after liver transplantation, indicating this pathway plays an important role in the occurrence of ACR.

Objective To characterize emotional memory impairment in untreated patients with Parkinson' s disease (PD) in the early stages of the disease. Methods Emotional memory tasks using standardized sets of emotional pictures including positive, neutral and negative valence were tested in 33 untreated patients with PD in the early stages and 31 healthy controls matched with age, sex, and education.Results Compared with the healthy adults (13.4 ± 1.4), PD patients had significant loss of negative valence picture memorizing ( 8. 9 ± 1.0; t = - 14. 87, P ＜ 0. 01 ).There was no significant difference between PD patients and normal controls in positive ( 11.8 ± 1.0 vs 12. 4 ± 2. 2 , t = - 0. 95 ) and neutral (7.9 ± 1.4 vs 8.2 ± 1.3, t = - 0. 89) valence picture memorizing ( both P ＞ 0. 05 ). ConclusionThe emotional memory for negative valence pictures is impaired but the emotional memory for positive and neutral is relatively spared in early PD patients without treatment.

Objective To observe the effects of treadmill training and music therapy on freezing of gait among patients with Parkinson′s disease. Methods Fifty-six Parkinson′s disease patients with freezing of gait were randomly divided into a control group, a treadmill training group, a music group and a united group, each of fourteen. All groups received standard medication. The treadmill training group and the music group were also given treadmill training and music therapy respectively. The united group received both treadmill training and music therapy. All of the patients were assessed in terms of freezing score, time to start a first step, incidence of falls, UPDRS-Ⅲ scores and MBI scores before and after 8 weeks of intervention. Results There were no sig-nificant differences among the 4 groups before the training. Significant differences were observed in all of the measurements between the treadmill training and united groups after the treatment. In the music group, only the average UPDRS-Ⅲ and MBI results were significantly improved after the treatment. Compared with the treadmill training and music groups, the united group showed significant differences in all of the measurements. Conclu-sion Treadmill training combined with music therapy is more effective than either alone in relieving gait freezing among those with Parkinson′s disease. It decreases falling and improves their capacity in the activities of daily living life, thus their quality of life.

Objective To study the role of vascular resection in the radical resection of hilar cholangiocarcinoma. Methods One hundred and forty-two cases of hilar cholangiocarcinoma (HC) data during July 2009 and December 2014 were analyzed retrospectively .After sufficient preoperative management including relieving jaundice , three dimensional assessments were evaluated and then surgical planning was designed. Results One hundred and forty-two cases underwent surgery after adequate preoperative preparation and evaluation, of whom 16 patients had vascular resection. Portal vein resection was performed in 11 cases, in which the portal vein wedge excision was performed in 4 cases, the portal vein anastomosis in 6 cases, and right anterior portal vein resection and reconstruction was performed in 1 case. Because of tumor invasion in the right hepatic artery, 4 patients underwent right hepatic artery resection and saphenous vein reconstruction, and 1 patient underwent right hepatic artery anastomosis. Postoperative complications were as follows:bile leakage in 1 case, no abdominal bleeding and abdominal infection, and no vascular thrombosis-related complications, and the overall complication rate was 1/16. There was no perioperative death . The follow-up results was as follows: 1 patient died 3 months after surgery, 1 died of liver metastasis after 9 months, 2 cases were lost to follow up, and the rest patients were still followed up. Conclusions Vascular resection for hilar cholangiocarcinoma helps to improve tumor resection rate and prognosis in large medical centers. With the use of three dimensional assessment, preoperative management and reasonable surgical strategy can reduce postoperative complications and ensure safety for hilar cholangiocarcinoma resection.

Objective To summarize the clinical characteristics,diagnosis and treatment of autoimmune pancreatitis(AIP).Methods The retrospective descriptive study was adopted.The clinical data of 43 patients with AIP who were admitted to the First Affiliated Hospital of China Medical University between January 2010 and December 2014 were collected.All patients underwent laboratory tests and imaging examinations.For the patients who had the definite diagnosis of AIP,corticosteroid therapy was adopted,and for the patients who had the suspicion of pancreatic cancer,surgical operation was carried out.Observed indexes included clinical manifestations,laboratory findings,imaging findings,treatment,pathological findings and follow-up status.The follow-up of imaging examinations and laboratory tests via outpatient and inpatient examinations was performed on the patients until January 2016.Measurement data with skewed distribution were presented as average (range).Results (1) Clinical manifestations:of the 43 patients,33 demonstrated obstructive jaundice,19 had loss of weight,18 had upper abdominal pain and 5 had acute mild pancreatitis (the same patient with a variety of symptoms).Extrapancreatic organ involvement was detected in 23 of the 43 patients,including focal nephritis in 15 patients,sicca syndrome in 7 patients,pulmonary involvement in 5 patients,autoimmune thyroiditis in 5patients,hilar IgG4-associated cholangitis in 3 patients,retroperitoneal fibrosis in 3 patients and ulcerative colitis in 1 patient (the same patient with a variety of diseases).(2) Results of laboratory tests:17 patients underwent serum IgG4 detection and increased expression of IgG4 was found in 15 patients.(3) Imaging findings:all the 43patients underwent enhanced computed tomography (CT) scan of pancreas.Diffused enlargement of pancreas with appearance of sausage was observed in 34 patients and localized pancreatic head enlargement was observed in 9patients.Twenty patients had peripancreatic vascular involvement including stenosis or occlusion of splenic vein,stricture at the confluence of superior mesenteric vein,portal vein and splenic vein resulting in regional portal hypertension which led to gastric venous varices and splenomegaly.Three patients showed that superior mesenteric artery was surrounded by tissue masses with massive ascites.Five patients had pancreatic pseudocyst with splenic vein involvement and peripancreatic effusion.The abdominal CT examination of 15 patients with renal involvement showed renal cortical wedge or nodular lesions.Pulmonary inflammatory pseudotumor and interstitial pneumonia appeared in 5 patients,CT of whom showed solid pulmonary nodules or diffuse ground-glass opacities and patchy shadows in pulmonary interstitium.Thirty-two patients underwent magnetic resonance cholangiopancreatography and 28 patients underwent endoscopic retrograde cholangiopancreatography.The results showed the focal and diffuse or segmental stenosis of the main pancreatic duct were detected in 28 patients (diffuse stenosis in 23patients,local stenosis in 5 patients),stenosis of the intrapancreatic common bile duct in 32 patients,stenosis of hilar bile duct in 3 patients.Nine patients underwent endoscopic ultrasonography examination.The results showed 3 patients had hypoechoic mass in the head of pancreas and 6 patients had diffuse or heterogeneous hypoechoic of the pancreatic parenchyma.(4) Treatment status:of 43 patients,30 patients received initial steroid treatment.After steroid treatment,the clinical symptoms of 27 patients gradually improved and laboratory and imaging findings were both improved.Eleven patients didn 't receive initial steroid treatment,of whom 2 patients with obstructive jaundice underwent endoscopic biliary stent implantation,7 patients underwent surgical biliary drainage and 2 received neither endoscopic treatment nor surgical treatment.Two patients underwent pancreatoduodenectomy due to suspicion of pancreatic adenocarcinoma and were diagnosed as AIP based on postoperative pathological results and treated with steroid after operation.(5) Pathological examination:of the 43 patients,22 underwent pathological examination.Results of pathological examination of superficial lymph nodes showed lymph follicles hyperplasia,and immunohistochemistry staining showed massive IgG4 positive plasmocytes in 5 patients,supporting the diagnosis.Results of fine needle aspiration cytology in 15 patients showed no cytological evidence were detected in 14 patients and epithelial cells of malignant tumors in 1 patient.Results of pathological examination of surgical specimens showed dense infiltration of lymphocyte and immunohistochemistry staining showed massive IgG4 positive plasmocytes.(6) Follow-up status:of 43 patients,41 were followed up for an average time of 28 months (range,12-71 months).Of the 30 patients who received steroid treatment,27 had complete response and 3 had partial response.Of the 27 with complete response,4 patients relapsed during followup and were cured by continuous steroid treatment and extending treatment course,3 patients did not relapse,1 patient relapsed again,and long-term low dose steroid maintenance therapy was needed.The 3 patients with partial response were treated with mycophenolate mofetil for 6,4,4 months and imaging abmormalities disappeaved absolutely.Eleven patients received no steroid for the initial treatment,in whom 2 patients were lost to follow-up,1 patient with common bile duct jejunum anastomosis died in 1 year after surgery.In the other 8 patients,the imaging abnormalities were spontaneous remission or clinical and imaging manifestations were improved after steroid treatment and no recurrence was found.Two patients who underwent pancreatoduodenectomy due to suspicion of pancreatic adenocarcinoma received steroid treatment after operation and no recurrence was observed.Conclusion Clinicians should combine clinical manifestations,serological examination,imaging findings and pathological examination to make definite diagnosis of AIP and make differential diagnosis of pancreatic cancer to avoid unnecessary surgical treatment by misdiagnosis.

BACKGROUND:Although vitamin C has an anti-oxidation role and can promote cell proliferation, there is a lack of research about the promoting effect of vitamin C on the proliferation of adipose-derived stem cells under high glucose conditions and the related molecular mechanisms.OBJECTIVE:To explore the promoting effect of vitamin C on the proliferation adipose-derived stem cells treated by the high glucose and the related molecular mechanisms.METHODS:Passage 3 human adipose-derived stem cells were cultured under high glucose conditions and then treated with different concentrations of vitamin C (0, 100, 150, 200, 250, 300 μmol/L). Cells cultured under low glucose conditions acted as controls. The expression levels of p-ERK and p-AKT proteins were detected by western blot. MTT method was used to choose the optimal concentration and time of vitamin C for all the subsequent tests. Human adipose-derived stem cells cultured under high glucose conditions were divided into four groups, and cells in blank control group had no treatment. Cells in the other three groups were treated with the optimal concentration of vitamin C (vitamin C group), LY294002+the optimal concentration of vitamin C (LY294002 group), or U0126+the optimal concentration of vitamin C (U0126 group) for 48 hours.EdU staining assay was used to detect the cell proliferation of human adipose-derived stem cells.RESULTS AND CONCLUSION:(1) Cell counting kit detection:We found that high glucose reduced the proliferation of human adipose-derived stem cells, and vitamin C promoted the proliferation of these cells. The best concentration of vitamin C was 200 μmol/L and the optimal effect time was 48 hours. (2) Western blot detection:Compared with the 0 μmol/L vitamin C group, the level of p-ERK in the 200 μmol/L vitamin C group was upregulated significantly (P < 0.01),while no significant expression change in p-AKT protein was found in control, 0 and 200 μmol/L vitamin C groups.(3) EdU test:the number of EdU positive cells was significantly higher in the vitamin C, LY294002, and control groups compared with the blank control group (P < 0.01). Moreover, compared with the vitamin C group, the EdU positive cells in the U0126 group were decreased significantly in number (P < 0.01). In conclusion, the ERK/MAPK signaling pathway is involved in the promotion effect of vitamin C on the proliferation of human adipose-derived stem cells under high glucose conditions.

Objective To assess the characteristics and influencing factors of non-motor symptoms (non-motor symptoms) in untreated patients with Parkinson's disease (PD) at early stage.Methods A total of 106 untreated patients with PD at early stage and 108 healthy controls (HC) matched for age,gender,education status and occupation were investigated by non-motor symptoms questionnaire.And both of them were divided into two groups:aged group (≥60 years old) and nonaged group (＜60 years old).76 cases with an average of (66.8±6.6) years old were selected in the aged group,and 30 cases with an average of (57.3± 5.2) years old were selected in the non-aged group.The distribution of non-motor symptoms items was analyzed between two groups.The clinical characteristics of PD patients were also collected to assess the dominant influencing factors for the prevalence of non-motor symptoms.Results The total number of non-motor symptoms was significantly higher in aged PD patients and non aged PD patients than in age-matched control groups [(13±7) vs.(4±2),t=10.843; (11±4) vs.(2±1),t=12.145,both P＜0.001].The incidence of non-motor symptoms was significantly higher in aged PD patients (20 items) and non-aged PD patients (9 items) than in age-matched control groups (both P＜0.05).The constipation in aged PD patients and the olfactory disturbance in non-aged PD patients were the most frequent ones.The number of non motor symptoms was positively correlated with course of disease(r=0.413),age(r=0.385) and Hoehn-Yahr stage (r =0.318) in aged PD patients (all P ＜ 0.05).The number of non motor symptoms was positively correlated with Hoehn-Yahr stage in non-aged PD patients (r=0.306,P＜0.05).Conclusions non-motor symptoms are more frequently seen in aged and non-aged PD patients than in age-matched controls.non-motor symptoms are more common in PD patients at earl stage and the number of non-motor symptoms increases with the degree of disease severity and age.More attention should be paid to the diagnosis and rational treatment of the non-motor symptoms so as to increase treatment effectiveness.

Objective To observe the effects of levodopa on verbal and spatial working memory in elderly patients with Parkinson's disease (PD).Methods The modified Smith working memory software were applied to study the verbal and spatial working memory in 32 PD patients before and 8 weeks after treatment with levodopa and benserazide hydrochloride tablets.The results of PD patients were compared with those in 32 normal controls matched for age,sex and education.Results The correct rates of phonological verbal and coordinate visuospatial working memory were significantly lower in the PD patients without treatment than in the normal controls (both P＜0.01).The correct rates of semantic verbal and categorical visuospatial working memory had no significant differences between the PD patients without treatment and the normal controls (both P＞0.05).The correct rates of phonological verbal working memory and coordinate visuospatial working memory were significantly higher in the PD patients after administration of levodopa and benserazide tablets than in PD patients without treatment [(88.6±6.5)％ vs.(82.1 ± 6.6)％,(84.2±8.8)％ vs.(75.2±8.9)％,respectively,both P＜0.01],but there were no significant differences in the correct rates of semantic verbal working memory and categorical visuospatial working memory between the above two groups (both P＞0.05).The correct rates of phonological verbal,semantic verbal,coordinate visuospatial and categorical visuospatial working memory were negatively correlated with the course of disease,and had no correlations with age and the degree of education.Conclusions Levodopa can improve the functions of phonological verbal working memory and the coordinate visuospatial working memory,but has no effect on the semantic verbal working memory and the categorical visuospatial working memory.It is postulated that dopamine replacement therapy may have double dissociable effects on the different working memories.

Objective To elucidate the resistance mechanisms of clinical colistin-resistant Klebsiella pneumoniae and Escherichia coli isolates in China.Methods A total of 964 K.pneumoniae and 1 389 E. coli isolates were retrospectively collected from national surveillance programs from 2011 to 2014 in China. Antimicrobial susceptibility testing was determined by the microdilution method.The PCR amplification followed by sequencing was used to detect the mcr-1 gene and colistin-resistance genes, including mgrB, pmrB and phoQ.Real-time quantitative PCR was performed to examine the relative transcriptional levels of pmrB, pmrC, pmrD, pmrK and pmrE genes in K.pneumoniae, and pmrA, pmrB, pmrC, phoP and phoQ genes in E.coli.Conjugation experiment was used to detect the transferability of the resistance plasmid carrying the mcr-1 gene.Statistical analyses were performed using IBM SPSS Statistics (version 16.0) and a P value <0.05 was considered statistically significant. Results The colistin-resistant rates of K. pneumoniae and E.coli were 0.62% ( 6/964 ) and 1.66% ( 23/1 389 ) , respectively.No amino acids substitutions were identified in mgrB genes among colistin-resistant isolates.Among six colistin-resistant K. pneumoniae isolates, five isolates were identified to have point mutations in pmrB gene, but no point substitution was detected in phoQ gene.One to four point mutations had been found in pmrB and phoQ genes in colistin-resistant E.coli isolates, respectively.The expression level of pmrB, pmrC, pmrD, pmrK and pmrE genes showed no significant difference between colistin-resistant and colistin-susceptible isolates [pmrB, (1.04 ±1.12) vs.(0.94 ±0.67), P=0.945; pmrC, (1.39 ±2.01) vs.(0.16 ±0.27), P=0.101;pmrD, (1.59 ±2.43) vs.(0.88 ±0.34),P=0.445;pmrK, (0.64 ±0.62) vs.(0.04 ±0.10), P=0.051;pmrE, (3 492 833 388.83 ±8 478 977 986.85) vs.(20 771 428.93 ±38 000 732.85), P=0.445].However, the transcriptional level of pmrB genes in colistin-resistant group was 9.5-fold higher than that of the colistin-susceptible group in E.coli isolates.Four in six colistin-resistant K.pneumoniae isolates possessed mcr-1 gene, whereas all of the colistin-resistant E. coli had the mcr-1 gene. The conjugation verified the transferability rate of the plasmid carrying mcr-1 gene was 5.78 ×10-6 , and the MIC value of colistin of the conjugant increased 21-fold than the recipient strain.Conclusions Plasmid-mediated mcr-1 gene was the major reason for colistin resistance in clinical isolates of K.pneumoniae and E.coli. Some other resistance mechanisms such as transcriptional up-regulated pmrB gene also involved in colistin resistance.

Objective To investigate the effect of nerve growth factor ( NGF) combined with exercise train-ing on nerve protection, learning and memory. Methods Twenty-four six-month-old APP/PS1 mice were randomly divided into a control group, an NGF group, an exercise group and an NGF combined with exercise training group ( the combined group) , each of 6. The control group mice were given 50 μL of saline intra-nasally every day, while the NGF group were given 50 μL of NGF solution (0.1 mg/ml) intranasally. The exercise group was given 50μL of saline intra-nasally plus 30 minutes of treadmill exercise daily, while the combined group was given 50μL of the NGF solution and 30 minutes of exercise. The intervention lasted 21 days. Then Morris water maze test was conducted to e-valuate the memory of all groups, and all of the mice were sacrificed and immunohistochemical staining was used to check the levels of APP, Ki-67 expression in cortex and NeuN expression in hippocampus. Results In the place navigation test, significant differences in escape latency were observed among the 4 groups, with the NGF, exercise and combined groups showing average latencies significantly shorter than that of the control group The average latency of the combined group was also significantly shorter than that of the NGF and exercise groups. In the spatial probe, there were significant differences among the 4 groups in the percentage of time spent in the target quadrant. The com-bined group′s average was significantly larger than those of the NGF and exercises groups. The immunochemistry showed that the combined group had better-arranged neurons and more cells compared with the 3 other groups. The average APP protein expression of the combined group was significantly lower than that of the other groups, but their Ki-67-positive and NeuN-positive cells were found to be significantly more numerous. Conclusion NGF combined with exercise can improve learning and memory, at least in APP/ PS1 mice. This might be associated with its reduc-ing the expression of APP and promoting the expression of Ki-67 and NeuN.