Objective:To explore the influential factors of cognitive change among Chinese oldest-old.Method: Three waves of data from the Chinese Longitudinal Healthy Longevity Survey(CLHLS)were analyzed with a two-level repeated measures model.Results:In baseline,the male had a higher mean MMSE score than the female(27.0?3.7/24.4?5.6,P

OBJECTIVETo compare the tissue engineered cartilage constructed with chondrocytes derived from auricular and articular cartilage.

METHODSChondrocytes were isolated from porcine auricular and articular cartilage, and BMSCs were obtained from bone marrow aspirate and cultured. Each kind of chondrocytes were resuspended alone or mixed with BMSCs at a ratio of 1:1, and seeded onto PGA/PLA scaffolds to construct tissue engineered cartilage (n = 4). The constructs were cultured for 8 weeks in vitro and then subcutaneously implanted into nude mice for 6 weeks. The differences between chondrocytes monoculture from articular and auricular cartilage or between each of them co-cultured with BMSCs were evaluated by gross view, measurement of thickness and wet weight, histological examinations including H&E, Safranin O, type II collagen, and Ponceau's & Victoria blue staining, and gene expression analysis of cartilage related genes.

RESULTSNo obvious differences were found histologically among the complexes constructed in vitro 8 weeks except for few elastic fibers secreted in the auricular chondrocytes + BMSCs co-culture group. Neo-cartilage is thicker in the groups of articular chondrocytes (38. 1% than the group of auricular chondrocytes, P < 0.05) and articular chondrocytes + BMSCs co-culture (19.3% than the group of auricular chondrocytes + BMSCs, P < 0.05). However, after 6 weeks in vivo the elastic fibers were found positive in the complexes constructed by auricular chondrocytes, and its staining was even stronger and more homogenous in the group of auricular chondrocytes + BMSCs co-culture. The tissues generated by articular chondrocytes alone and co-cultured with BMSCs both formed the characteristic features of three-layer structure of hyaline cartilage and ossified in vivo with significant up-regulation of COL10A1 and MMP-13. To summarize, auricular chondrocytes formed the elastic cartilage while articular chondrocytes formed the hyaline cartilage during the development of tissue engineered cartilage either by monoculture or the co-culture with BMSCs.

CONCLUSIONSThe chondrogenic response of chondrocytes from different cartilage origins demonstrates that an initial chondrocyte and cartilage type recapitulates the same in later tissue-engineered development.

Animals ; Bone Marrow Cells ; cytology ; Cartilage, Articular ; cytology ; Cells, Cultured ; Chondrocytes ; Coculture Techniques ; Ear Auricle ; cytology ; Mesenchymal Stromal Cells ; cytology ; Mice, Nude ; Swine ; Tissue Engineering ; methods ; Tissue Scaffolds

Objective To observe the effectiveness of introducing Diazepam into combined aesthesia of Sumianxin and ketamine hydrochloride .Method A total of 80 rabbits of both genders for operation were randomly divided into A , B and C groups .The A group was injected with Sumianxin intramuscularly ( 0.3 mL/kg by weight ) .The B group was injected with Sumianxin and ketamine hydrochloride intramuscularly ( 0.3 mL/kg by weight ) .The C group was injected with Diazepam intravenously ( 1.5 mL/kg by weight ) combined with Sumianxin and ketamine hydrochloride injected intramuscularly (0.3 mL/kg by weight).The aesthetic effects, induction time, anesthesia maintaining time, total anaesthetic dose and operation time were observed , recorded and compared .Result The induction time of the C group was significantly shorter than A and B groups (P<0.01).The initial anesthesia maintaining time of the C group was the longest among the three (P<0.01) with least total anaesthetic dose (P<0.01).The operation time of the C group was the least with best aesthetic effects (P<0.01).Conclusion Introducing Diazepam into combined aesthesia of Sumianxin and ketamine hydrochloride can improve the aesthetic effects .Therefore , this is an optional aesthetic method for time-consuming animal operation or sensitive surgical sites of rabbits .

AIMTo research the spontaneous firing activities during different-frequency stimulation of subthalamic nucleus and microelectrophoresis GABA, Glu and their antagons respectively, approaching the mechanism of DBS in the treatment of Parkinson's disease further.

METHODSUsing extracellular recording to investigate the effect of different-frequency stimulation of STN and microelectrophoresis several drugs on the spontaneous firing activities of the SNr neurons.

RESULTSFor STN stimulation at low frequency, there was no difference on the spontaneous firing activities of SNr neurons between pro-stimulation and meta-stimulation (P > 0.05). With the increasing of stimulation frequency, most of the SNr neurons were inhibited. While during the STN stimulation frequency at high-frequency, the firing rates of inhibited SNr neurons were changed (P < 0.05). Glu had catatonic excitement effect on the SNr neurons, whereas GABA had tonic inhibition effect. 80% of SNr neurons which were inhibited by STN-HFS were not inhibited by STN-HFS on the basis of excitatory effect of BIC.

CONCLUSIONTo treat the motor symptoms of PD, when SIN is selected as the target nucleus, the electrical stimulation with high-frequency should be chosen. It is possible that SIN-HFS modulate the activity of SNr by inhibitory effect of GABA predominantly.

Action Potentials ; physiology ; Animals ; Electric Stimulation ; Electrophoresis ; methods ; Glutamic Acid ; administration & dosage ; pharmacology ; Male ; Neurons ; physiology ; Parkinson Disease ; physiopathology ; therapy ; Rats ; Rats, Sprague-Dawley ; Substantia Nigra ; physiology ; Subthalamic Nucleus ; physiology ; gamma-Aminobutyric Acid ; administration & dosage ; pharmacology

OBJECTIVETo investigate the effect of allogene hepatic nonparenchymal cell (NPC) on the survival of grafted skin in mice and its underlying mechanism.

METHODSSixty-five C(3)H and fifty-eight C(57)BL/6 mice were employed in the study. Twenty C(3)H mice were used as skin donor and forty as the source of hepatic NPC. The rest five served as the stimulators of mixed lymphocyte culture (MLC) before and on the 7th, 18th, 30th, and 60th day after NPC infusion with 1 at each time point. MLC was determined and expressed as count per minute (CPM). Fifty-eight C(57)BL/6 mice were further divided into experimental (E, n = 50) and control groups (C, n = 8). The mice in C group only underwent skin grafting without NPC infusion. The mice in E group received with 2 x 10(7) NPC via caudal vein, followed by peritoneal injection of cytoxan (200 mg/kg) 48 hours later; They were grafted with skin donated from C(3)H mice 18 days after injections. The survival time of the mice in the two groups was observed. The serum levels of interleukin-4, chimera and MLC in the two groups were determined before and on 7th, 18th, 30th, 60th days after NPC infusion, and micro-chimera were aslo assessed on the 1st and 3rd day after NPC infusion. Five mice were sacrificed at each time point.

RESULTSThe survival time of skin graft in E group (70.0 +/- 17.2 day) was obviously longer than that in C group. The serum levels of IL-4, chimera in E group were increased gradually, while MLC response decreased gradually. The serum IL-4 level reached 251.5 +/- 11.0 ng/L and splenic chimera level to 26.30 +/- 1.04% on the 60th day after NPC infusion.

CONCLUSIONThe high levels of IL-4 and chimera might play important roles in inducing and maintaining immune tolerance.

Animals ; Female ; Graft Survival ; immunology ; Hepatocytes ; immunology ; Immune Tolerance ; Interleukin-4 ; metabolism ; Male ; Mice ; Mice, Inbred C3H ; Mice, Inbred C57BL ; Skin Transplantation ; immunology ; Surgical Flaps ; Transplantation Chimera ; Transplantation, Homologous ; immunology

AIMTo approach the protective effect of low dose gentamicin against high ototoxic dose of gentamicin.

METHODSThe guinea pigs were randomly divided into four groups: control group, low dose group, low dose protective group and high dose group. Each group received multiple intraperitoneal injections of gentamicin sulphate within different durations. Auditory brain stem response (ABR) was examined one day previous to the first and 24 h after the final injection respectively. The bulla was taken out so that the content of NO, MDA and the activity of LDH in cochlear were determined.

RESULTSThe threshold of ABR was significantly lower in low dose protective group compared with high dose group (P < 0.01). The content of NO (15.86 +/- 1.98 nmol/mg pro) and MDA (19.14 +/- 0.96 nmol/mg pro) in homogenate of high dose group was significantly higher than that of control group, low does group and low does protective group (P < 0.01). The increase of the content of NO and MDA induced by high dose GM could be significantly decreased by low dose GM administration previous to high dose injection (P < 0.01). The activity of LDH in homogenate of high dose group was significantly higher compared with control group, low dos group and low dos protective group (P < 0.01). There was no statistically significant difference of content of NO and MDA among control group, low does group and low does protective group.

CONCLUSIONThe protective effects resulting from previous low dose administration to high dose injection of GM may be related to the decrease of content of NO and MDA and activity of LDH both of which induced by high dose GM.

Animals ; Cochlea ; metabolism ; Evoked Potentials, Auditory, Brain Stem ; physiology ; Female ; Gentamicins ; administration & dosage ; adverse effects ; Guinea Pigs ; Hearing Loss ; chemically induced ; prevention & control ; Male ; Malondialdehyde ; metabolism ; Nitric Oxide ; metabolism

ObjectiveTo investigate the clinical features and risk factors for patients with liver failure complicated by invasive pulmonary aspergillosis (IPA), and to provide a reference for clinical diagnosis and treatment. MethodsThe clinical data of 477 patients with liver failure who were diagnosed and treated in Henan Provincial People′s Hospital from January 2010 to December 2014 were collected, and the clinical features, laboratory markers, and results of imaging examinations of patients with IPA were retrospectively analyzed. Another 49 patients with liver failure who were hospitalized within the same period, had similar ages, and were not complicated by pulmonary infection were randomly selected as controls. The independent samples t-test was used for comparison of continuous data between groups, the chi-square test or Fisher′s exact test were used for comparison of categorical data between groups, and multivariate logistic regression analysis was performed to analyze the risk factors for liver failure complicated by IPA. ResultsAmong the 447 patients with liver failure, 43(96%) were complicated by IPA. Age (P=0.023), hepatic encephalopathy (P=0.021), long-term use of broad-spectrum antibiotics (P=0.007), use of hormone (P=0.016), and deep venous catheterization (P＜0.001) were independent risk factors for the development of IPA. Clinical manifestations of liver failure patients with IPA lacked specificity. Lung CT scan showed multiple nodules, masses, and wedge-shaped consolidation near the pleura in both lungs, but typical halo sign and air crescent sign were rarely seen. Among the 35 patients who received antifungal therapy, 30 were improved or cured, 3 died of digestive tract bleeding, 2 clied of plumonary infection, and all the other patients who did not receive therapy also died. ConclusionPatients with liver failure have various risk factors for the development of IPA, and the clinical manifestations are not typical, with high incidence and fatality rates. Early detection and treatment is the key to improving survival rates.

Aim To investigate the effect of thioester-ase superfamily member 4(THEM4)expression on col-lagen secretion in human renal proximal tubular epithe-lial cells (HKC)treated with high glucose. Methods In order to examine the direct effect of THEM4 ex-pression vector on PI3K/ Akt pathway and collagen se-cretion,pYr-ads-4-THEM4 expression vector was con-structed and transfected into the HKC with lipo-fectamine 2000 in vitro. HKC cells were randomly di-vided into four groups:normal glucose group (Con-trol),high glucose group (HG),high glucose plus pYr-ads-4-THEM4 vector group (HG + THEM4 vec-tor) and high glucose plus pYr-adshuttle-4 vector group (HG + V vector). After 48 h with HG stimula-tion,the cells were collected for extraction of protein and phospho-Akt (Ser 473),THEM4,TGF-β1 andα-SMA protein expression were examined by Western blot and immunofluorescence staining respectively. Col Ⅰ and Col Ⅲ were detected using the competitive sandwich ELISA kit according to the manufacturer's instructions. Results High glucose inhibited THEM4 expression,and induced increased phospho-Akt (Ser 473),TGF-β1,α-SMA and secreted ColⅠand secre-ted Col Ⅲ in HKC cells. Up-regulation of THEM4 re-versed high glucose-induced decreased THEM4,in-creased phospho-Akt (Ser 473),TGF-β1,α-SMA, secreted Col Ⅰ and secreted Col Ⅲ in HKC cells. Conclusion The up-regulation of THEM4 may de-crease Col Ⅰ and Col Ⅲ secretion by inhibiting the phosphorylation of Akt and down-regulating the expres-sion of TGF-β1 and α-SMA in high glucose-induced HKC cells.

Purpose To investigate the relations of THEM4/Akt expression and extracellular matrix deposit in kidney of diabetic mice. Methods Diabetic mice models were successfully established by intraperitoneally injected STZ. Both normal control mice and diabetic mice were raised for 8 week until they were sacrificed. Western blot, immunohistochemistry and realtime PCR were used to detect the expression of THEM4, phospho-Akt (Ser 473), TGF-β1, a-SMA, Col Ш, FN protein and THEM4 mRNA in the kidneys of normal mice and diabetic mice. Results Compared to normal control mice, THEM4 expression decreased by 37.7％ followed by 3.66, 1.29 2.33, 1.99 and 2.82 times increased of phospho-Akt (Ser473), TGF-β1, a-SMA, Col Ш and FN in kidney of diabetes mellitus. Extracellular matrix accumulation was found in renal interstitial region of diabetic mice. Conclusion The decreased THEM4 might cause extracellular matrix deposit in kidney of diabetic mice by upregulating the phosphorylation of Akt and TGF-β1, α-SMA expression in diabetic mice.

BACKGROUNDThe extended spectrum β-lactamase (ESBL)-producing Escherichia coli (E. coli) and Klebsiella pneumoniae (K. pneumoniae) are the major pathogens causing pneumonia and have a significant impact on the clinical course. Limited data exist on molecular characterization of ESBL-producing E. coli and K. pneumoniae that cause pneumonia. The aim of this study was to investigate the comprehensive multilevel characteristics of E. coli and K. pneumoniae causing pneumonia in China for the first time.

METHODSE. coli (17) and K. pneumoniae (21) isolates responsible for pneumonia were isolated from 1270 specimens collected in a prospective multi-center study in eight teaching hospitals in China from June to December in 2007. The susceptibilities, ESBL confirmation, sequence typing, blaCTX-M and blaSHV genes, their genetic environment and plasmid Inc/rep types were determined.

RESULTSSixteen E. coli (94.1%) and eleven K. pneumoniae (52.4%) isolates were ESBL producers. About 77.8% and 66.7% of them were resistance to ciprofloxacin and levofloxacin, and 100% were susceptible to imipenem. The most prevalent ESBL gene was CTX-M-14, followed by SHV-2, CTX-M-15, CTX-M-3, CTX-M-65, SHV-12, SHV-26 and SHV-28. SHV-1 and SHV-11 were also detected and coexisted with blaCTX-Ms in five strains, and three strains contained only SHV-1. All CTX-M-14 were detected ISEcp1 upstream and nine were found IS903 downstream and the majority of them (64.3%) were carried by IncF plasmids. All blaSHV were flanked by recF and deoR, located on IncF, IncN, IncX and IncH plasmids. Two SHV-2, one SHV-1 and the only SHV-28 were further preceded by IS26. Genes lacY and lacZ were detected at further upstream of two blaSHV-1. The K. pneumoniae carrying SHV-28 was susceptible to β-lactams, and no mutations or deletions in gene or promoter sequences were identified to account for susceptibility. Multilocus sequence typing experiments showed the ESBL-producing strains were genetically diverse.

CONCLUSIONSThe rate of occurrence of blaESBL in E. coli and K. pneumoniae causing pneumonia was high, and blaCTX-M-14 was dominant and probably mobilized by ISEcp1 mainly on IncF plasmids. Importantly, unexpressed blaESBL genes may occur in susceptible isolates and hence may have clinical implications.

Blotting, Southern ; Escherichia coli ; enzymology ; genetics ; Klebsiella pneumoniae ; enzymology ; genetics ; Plasmids ; genetics ; Pneumonia ; microbiology ; Promoter Regions, Genetic ; genetics ; Prospective Studies ; beta-Lactams ; metabolism