Objective To study the value of relative cerebral blood flow(rCBF)changes in patients with amnestic-type mild cognitive impairment (aMCI)and mild Alzheimer disease(AD) using MRI pulsed arterial spin labeling(PASL).Methods A prospective study recruited 37 aMCI patients (aMCI group),30 mild AD patients(mild AD group) and 30 healthy volunteers (normal control group) from March 2011 to December 2013,MRI using PASL for cerebral perfusion imaging was performed and data of rCBF were collected.Taking age as covariate,analysis of variance (ANONA)was carried out to assess the difference of rCBF among all the three groups,then Bonferroni was done between every two groups.A follow-up examination using PASL was performed in the seventeen patients of the aMCI group.And paired t-test was used for comparing the longitudinal change of their rCBF data.Results Compared with the normal control group,the aMCI group showed significant increase of rCBF in bilateral posterior cingulate cortices and precuneus (cluster number 2 785,P＜0.05).While the mild AD group showed decrease of rCBF in the left inferior and superior parietal lobes,the angular,middle frontal lobe,as well as the right superior temporal lobe (cluster number 3 459-5 206,P＜0.05).When compared with the aMCI group,the mild AD group showed regional hypoperfusion in bilateral middle frontal lobes,the left precuneus,the right postcentral and inferior parietal lobe (cluster number 3 236-19 863,P＜0.05).In the longitudinal study of the 17 aMCI patients,an increased rCBF was found to coexist with reduced rCBF in the left inferior frontal and lateral occipital cortex,bilateral frontal poles and paracingulate gyrus,with hyperperfusion dominated.Increased rCBF was also detected in the left temporal lobe,the angular gyrus and precuneus,while decreased rCBF was present in the left putamen,the operculum and right corpus callosum (P＜0.05).Conclusions ASL perfusion imaging is a valuable method for dynamic monitoring of the cerebral perfusion changes in aMCI and AD patients.PASL will assist in finding a useful imaging biomarker for early diagnosis of AD.

Objective To investigate the protective effect of agmatine (AGM) against peritoneal inflammatory response and neutrophil (PMN) infiltration induced by zymosan (ZYM) in mice. Methods Thirty-six adult male C57BL/6 mice were randomly divided into sham group, model group, and AGM treatment group. Peritonitis model was reproduced by intra-peritoneal injection of 1 mg/mL ZYM (0.5 mL), while equivalent phosphate buffer saline (PBS) was given to sham group. 200 mg/kg AGM was injected into peritoneal cavity after ZYM challenge in AGM treatment group. Six mice in each group were sacrificed at 2 hours and 6 hours, respectively, after reproduction of the model. Blood sample and peritoneal lavage fluid (PLF) were collected. The levels of keratinocyte-derived chemokine (KC), macrophage inflammatory protein 2 (MIP-2), tumor necrosis factor-α (TNF-α), interleukins-6 (IL-6) in serum and PLF were determined by enzyme linked immunosorbent assay (ELISA). The number of leukocytes and PMN in PLF were determined by hemocytometer and flow cytometry, respectively. Results Compared with sham group, all serum and PLF levels of KC, MIP-2, TNF-α and IL-6 were greatly elevated at 2 hours after ZYM injection in model group, while AGM treatment could dramatically reduce the levels of the above-mentioned cytokines in serum and PLF as compared with those of the model group [serum KC (ng/L): 990.7±137.9 vs. 2 053.2±262.7, MIP-2 (ng/L): 642.2±124.4 vs. 1 369.7±146.5, TNF-α (ng/L): 608.6±38.1 vs. 1 044.7±101.0, IL-6 (ng/L): 1 058.2±129.1 vs. 1 443.3±190.1; PLF KC (ng/L): 7 462.3±839.6 vs. 12 723.5±1 515.7, MIP-2 (ng/L): 1 570.8±193.4 vs. 3 471.4±384.7, TNF-α (ng/L): 1 115.8±156.7 vs. 1 499.2±231.2, IL-6 (ng/L): 2 646.5±223.2 vs. 3 126.7±291.4; all P < 0.05]. The expressions of KC, MIP-2 and TNF-α at 6 hours were significantly lower than those at 2 hours in model group and AGM treatment group, but IL-6 levels were further increased. The levels of KC and MIP-2 in serum and PLF at 6 hours were decreased to the levels of sham group. At 6 hours after the reproduction of the model, the number of total inflammatory cells and PMN of PLF in the model group was significantly higher than those of the sham group. In contrast, AGM notably lowered the number of inflammatory cells and PMN in peritoneal fluid after ZYM attack [total inflammatory cells (×109/L): 14.7±1.1 vs. 2.0±0.4, 10.1±1.2 vs. 14.7±1.1; PMN (×109/L): 11.37±1.22 vs. 0.18±0.05, 7.69±0.57 vs. 11.37±1.22, all P < 0.05]. Conclusion AGM can effectively alleviate acute peritoneal inflammatory injury induced by ZYM, mainly through reducing the secretion of inflammatory mediators and chemokines, and inhibiting the infiltration of leukocytes and neutrophils.

BACKGROUND: Thoracodorsal artery perforator flap can relieve damage to donor site and avoid bulk in the recipient site,but dissociation of perforating branch took time.Some one believed that it should be done by very experienced physicians and some muscle tissues should be reserved.OBJECTIVE: To investigate the method,effectiveness and clinical application of improved latissimus dorsi flap based on perforator flap conception for reconstruction of soft tissue defects of lower extremity.METHODS: A total of 17 patients needing skin flap transplantation were selected.12 latissimus dorsi musculocutaneous/muscle flaps,3 latissimus dorsi flaps with a few muscle and 2 double-leaf segmental latissimus dorsi compound flaps were designed based on perforator flap conception.According to the territory of latissimus dorsi musculocutaneous flap,a skin paddle in which anterior underlying muscle and main perforator was designed,extend about to the anterior edge of the latissimus dorsi muscle.An additional latissimus dorsi muscle flap was selected for soft tissue enlargement if necessary.Sometimes,double-leaf segmental latissimus dorsi musculocutaneous/muscle flap,including one muscle-sparing latissimus dorsi musculocutaneous flap and the other segmental latissimus dorsi muscle flap nourished by the lateral branches of the thoracodorsal vessels was selected to repair two adjacent defects.The harvested tissue area ranged from 12 cm×8 cm to 28 cm×17 cm.Survival state of skin flap,together with shape and function of donor site and recipient site of skin flap were observed.RESULTS AND CONCLUSION: Following skin flap transplantation,one case developed vascular crisis that was relieved following re-exploration for vessel anastomosis.All skin flap survived.Second-stage skin grafting was done on one muscle flap wound.All donor sites were sutured directly.After a follow-up of 3 to 18 months in 15 cases,only two cases received two-stage plastic operation because bulky flaps brought some trouble in wearing shoes.Improved latissimus dorsi flap based on perforator flap conception can reduce damage to the donor site and the receipt area bulk.Double-leaf segmental latissimus dorsi compound flaps can repair both heel and toe wound.The versatile latissimus dorsi flap designed using thoracodorsal artery perforator flap conception is an ideal flap for repairing widespread soft tissue defects in the lower extremity.

Objective To evaluate the clinical efficacy of small endoscopic sphincterotomy combined large-bal-loon dilation in treatment of common bile duct stones 1.0~2.5 cm in diameter. Methods 426 patients with large common bile duct (CBD) stones 1.0~2.5 cm in size were reviewed in our hospital between June 2010 and June 2014. They were randomized underwent small endoscopic sphincterotomy combined large-balloon dilation (SESPLBD) (n=218) or endoscopic sphincterotomy (EST) ( n= 208) for lithotripsy. The therapeutic outcome and complications were reviewed and compared. Results SESPLBD had higher complete duct clearance in one session (95.41 % vs. 93.75%), but there was no statistical significant difference. Bleeding was much less occurred in SESPLBD than in EST (2.29 % vs. 7.69 %, P= 0.025), especially when the stones were bigger than 1.5 cm in diameter. Mechanical lithotripsy was performed less in SESPLBD (13.76%vs 25.96 %, P=0.002), especially when the stones were 1.5 ~2.0 cm in diameter. There was no statistical significant difference in the incidence rate of post-ERCP pancreatitis (9.17 % vs. 6.73 %,P = 0.452), hyperamylasemia (19.72 % vs. 18.27 %,P = 0.796), perforation and death. Conclusions SESPLBD could be a safe method for large bile duct stones 1.0~2.5 cm in size. Compared with routine EST, it had less bleeding rate and mechanical lithotripsy requirement without increasing the incidence rate of post-ERCP pancreatitis or hyperamylasemia.

Objective To research the molecular biology characteristics and transient expression in BHK-21 cells of E geue segment from Japanese encephalitis virus(JEV) and construct an eukaryotic expression vector pEGFP-C1-JEV.Methods E gene segment of JEV was amplified by RT-PCR,construct the recombinant vector pEGFP-C1-JEV,which could express EGFP label proteins.Transfect pEGFP-C1-JEV vector into BHK-21 via LipofectAMINETM 2000,to observe expressing of EGFP label protein and transcription of aim gene,and to check up localization and antigenicity of expressed E protein by Immunohistochemistry and Western blot.Results It showed that the recombinant plasmid pEGFP-C1-JEV was successfully constructed and transfected to BHK-21 cells,the normal expression of green fluorescent protein expression rate was higher.RT-PCR showed that gene transcription in BHK-21 and normal expression,expression protein was mainly distributed in the cytoplasm of BHK-21 cells and the envelope in,and can with guinea pig anti-JEV antibody binding.Conclusion pEGFP-C1-JEV vector in BHK-21 cells was normal expression and there were no effect on cell growth and morphology.Meanwhile,on eukaryotic antigens was good antigenicity.This research as a base foundation for E protein gene of JEV eukaryotic expression and function in vitro and applied research.

ObjectiveTo analyze the characterstics of phenotype and genotype of multidrug resistant tuberculosis (MDR-TB) and extensively drug resistant tuberculosis (XDR-TB) by molecular line probe assay and liquid culture with MGIT960.MethodsGenoType MTBDR Kits were used for identifying the types of the first-line and second-line antituberculosis drug resistant genes partly and BD MGIT960 was used for detecting the chug susceptibility.Results( 1 ) Out of 94 MDR-TB strains,the rate of drug resistant to EMB,AMK,OFX and MFX by BD MGIT960 assay were 36.2%,17.0%,54.3% and 55.3%,respectively.Among these isolates,13 were extensively drug resistant tuberculosis (XDR-TB).(2) Compared with MGIT960,the concordance rate of GenoType MTBDRplus was 86.2% and 95.7% respectively.Taking MGIT960 results as reference,the sensitivity of GenoType MTBDRsl detecting the susceptibility of EMB,AMK,OFX and MFX to 94 isolates were 47.1%,81.3%,94.1%,94.2%,respectively.The specificity were 75.0%,98.7%,90.7%,92.9%,respectively.(3) Among the rpoB mutation categories,S531L accounts for most.MTB resistant to IFN caused by the mutation of katG chiefly and the S315T1 was in the majority.The gyrA mutation sites located at the ninety-fourth codon most.Out of 94 strains,23 were mixed with 2 kindsof Mycobacterium tuberculosis at least and 7 were undetectable mutations.Conclusion Among the M/XDR-TB,the strains resistant to INH,RFP,AMK,OFX and MFX were caused most by the mutation of katG,rpoB,rrs and gyrA,respectively.The relationship between EMB and embB was not so clear relatively.As a fast detecting drug susceptibility test kit,GenoType MTBDR possess good sensitivity and specificity.So,it could be as an assistant method to guide the therapy on clinic.

Objective To explore the effect of staff assistance program on clinical nurses' job burnout. Methods A staff assistance program was started and implemented by group counseling and individualized mental counseling for 104 clinical nurses. The job burnout scale was used to investigate their job burnout before and after the intervention. Result The score on"low personal achievement"score was higher than that before intervention, and the scores on"emotional failure"and"de-personalization"were lower than those before intervention (P<0.05). Conclusion The employee assistance program for clinical nurses can alleviate their job burnout.

Objective To explore Wang Xingkuan’s rules of syndrome and treatment of chest blocking and heartache (Xiongbixintong).Methods Collection of professor Wang Xingkuan’s 267 consilia of patients with Xiongbixintong for outpatients. Chinese medicine terminology was regulated and Excelldatabase was established. Symptom, syndrome element, pathogenesis and treatment were statistically described by using Weka3.6 software, and Apriori algorithm was adopted for the main pathogenesis→treatment analysis of association rules.Results Symptoms include:chest pain, heart palpitations, shortness of breath, pale tongue (dark) red, etc. Syndrome elements include:in liver, and heart, and blood stasis, phlegm, qi stagnation, etc. The key pathogenesis is liver-heart imbalance, including stagnation of liver qi, heart and blood stasis, deficiency of heart qi-ying, disturbing heart-mind, etc. The principle of treatment is liver-heart Tongzhi, so the treatment is of“liver” with Shu gan-mu;treatment of“heart” contains freeing channels, eliminating phlegm and blood stasis, quiet the heart, replenishing qi-ying, etc. The main pathogenesis related credibility→treatment was higher than 0.50;with high reliability, the liver-heart imbalance→liver-heart Tongzhi was 0.71. Medication includes catharsis and tonic,“catharsis” to salvia, allium macrostemon, pseudo-ginseng, bupleurum, etc;“tonic” to white ginseng, ophiopogon japonicus, radix paeoniae alba, poria with hostwood, polygala tenuifolia, etc. Conclusion “Xintongzhigan, liver-heart Tongzhi, catharsis and tonic” is Wang Xingkuan’s thoughts and experience in treating Xiongbixintong.

OBJECTIVETo study the mechanism underlying the effect of the SOCS3 rs4969170 A/G alleles on the occurrence of insulin resistance (IR) in patients with chronic hepatitis C.

METHODSThe promoter region of the SOCS3 gene was amplified by PCR,and luciferase expression vectors were constructed and transfected into HepG2,Huh7 cell lines.The relative luciferase activity of each expression vector was assessed by the dual luciferase reporter gene assay system.Western blotting was used to detect SOCS3 protein expression in PBMCs from groups of patients with the rs4969170 AA and AG genotypes.The state of IR in eight patients was evaluated by determining their HOMA-IR.

RESULTSThe pGL3-A, PGL3-G and pGL3-control vectors showed significantly different luciferase expression in the HepG2 cells (0.121 00 ± 0.022 07,0.027 00+/-0.012 49 and 0.043 33 ± 0.005 51; F =48.068, P=0.001) and in the Huh7 cell lines (0.164 70 ± 0.007 10,0.027 33 ± 0.017 04 and 0.033 67 ± 0.014 98; F =115.137, P=0.001). The expression of SOCS3 protein was significantly higher in the rs4969170 AA genotype group than in the AG genotype group (1.22 ± 0.40 vs. 0.30 ± 0.19; t =4.149, P=0.006).The IR index of patients with the rs4969170 AA genotype and the AG genotype was 4.11 ± 2.62 and 1.47 ± 1.01 respectively.There were three patients with IR in the rs4969170 AA genotype group and one in the rs4969170 AG group. There was no statistically significant difference between the two genotype groups (t=1.881, P=0.109).

CONCLUSIONSThe SOCS3 rs4969170 A haplotype may enhance transcriptional activity of the gene promoter to regulate gene expression, thereby increasing intracellular SOCS3 protein level and ultimately interfering with insulin signaling and causing IR in patients with chronic hepatitis C.

Cell Line, Tumor ; Genes, Reporter ; Genotype ; Haplotypes ; Hepatitis C, Chronic ; Humans ; Insulin Resistance ; Luciferases ; Polymorphism, Single Nucleotide ; Suppressor of Cytokine Signaling 3 Protein ; Suppressor of Cytokine Signaling Proteins

Objective:To investigate the mechanisms of effects of high fat and high fructose diet on rat aging.Methods:Adult male SD rats were divided into normal diet(ND) group and high fat and high fructose diet(HFHFD) group. After treatment for 48 weeks, these rats were sacrificed and the blood, liver, and brain tissues were collected. Serum triglyceride(TG), total cholesterol(TC), low density lipoprotein-cholesterol(LDL-C), high density lipoprotein-cholesterol(HDL-C) levels were determined by automatic biochemical analyzer. The serum levels of interluekin-2(IL-2), IL-6, and advanced glycation end products(AGEs) were measured using enzyme linked immunosorbent assay. The expressions of p16, p21, and p53 genes in the liver and brain tissues were detected by real-time quantitative PCR and western blot.Results:After 48 weeks treatment, there were significant differences in body weight and fasting plasma glucose between two groups. Serum TG, TC, and LDL-C in HFHFD group were significantly higher than those in the ND group( P<0.05), with an increase trend in HDL-C but without statistical difference. Compared with ND group, the level of IL-2 in HFHFD group was significantly decreased while the levels of IL-6 and AGEs were significantly increased(all P<0.05). The levels of p16 and p21 mRNA expressions as well as p53 and p21 protein expressions in liver and brain in HFHFD group were markedly increased compared with ND group(all P<0.05). Conclusion:Long-term high fat and high fructose diets accelerate the aging process of rats, which may be related to the damage of the immune system and the changes of cell senescence related gene expressions in liver and brain tissues.