OBJECTIVETo investigate the role of autophagy in acute lung injury (ALI) caused by multiple trauma in rats via pretreat with 3-methyladenine (3-MA).

METHODSForty-five Sprague-Dawley male rats, with age of 4 months and body weight of 250-300 g,were randomly divided into three groups. In the sham group, the rats received sphenotresia only;in the control group, the rats were made model of femur shaft fracture combined with brain injury, and treated with physiological saline by abdominal cavity at 1 hour before making model; in the 3-MA group, the rats were made model of femur shaft fracture combined with brain injury,and treated with 3-MA of 10 mg/kg by abdominal cavity at 1 hour before making model. Histologic changes and the concentration of related inflammatory factors in the damaged lung tissue were examined at 48 h after opteration, at the same time, the effect of 3-MA on the expression of LC-3 II and Beclin-1 was examined through reverse transcriptase polymerase chain reaction technique (RT-PCR).

RESULTSCompared with sham group, LC-3 II and Beclin-1 level in control group at 48 h after operation were obviously increased (P < 0.01). Compared with control group, LC-3 II and Beclin-1 level in 3-MA group at 48 h after operation were obviously decreased (P < 0.01). Compared with sham group, the level of proinflammatory cytokines (TNF-α and IL-6) in control group obviously enhanced (P < 0.01). Compared with control group, above items in 3-MA group was obviously lower (P < 0.01). Compared with control group,the histopathological damage of lung in 3-MA group obviously reduced (P < 0.01).

CONCLUSIONAutophagy can aggravate the acute lung injury caused by fracture of shaft of femur combined with brain injuries,but 3-MA can reduce tissue damage by inhibiting the autophagy and inflammatory response.

Acute Lung Injury ; prevention & control ; Adenine ; analogs & derivatives ; therapeutic use ; Animals ; Apoptosis Regulatory Proteins ; analysis ; Beclin-1 ; Interleukin-6 ; analysis ; Lung ; chemistry ; immunology ; pathology ; Male ; Multiple Trauma ; complications ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; analysis

Objective To study the significance and value of imaging typing by analyzing the correlation between contrast-enhanced ultrasound perfusion imaging typing and vascularization in patients with primary hepatocellular cancer.Methods The early enhanced arterial phase reflecting angioarchitecture,and the size and edge of enhanced tumor closely related to microvessel distribution, position and density, were observed and analyzed in 89 patients (113 lesions).All the cases were pathologically proved, and some received immunohistochemisty staining and the microvessel density were recorded.Results The perfusion imagings were classified into 4 types according to changes on contrast-enhanced ultrasound,dendritic,mixed,circular and net-like type.Atypia vessels were commonly seen in dendritic type,and Ⅲ and Ⅳ accounted for 79.1%.The microvessel density was higher than the other types.The circular type was relatively regular and Ⅲ and Ⅳ accounted for 12.5%.The microvessel density was lower than the other types.Combined with the contrast-enhanced ultrasound perfusion imaging and compared with pathological grading,growing methods and microvessel density, the dendritic type was characterized by infiltrative growth, and had strong invasion tendency.The circular type was characterized by expansive growth.And the other two types were characterized by transitional type.Conclusions Contrast-enhanced ultrasound can reflect the angioarchiteeture, the microvessel density and the position,and it is related to the pathological grading and growing methods.

Objective To investigate the plasma level change of myeloperoxidase(MPO)in patients suffering from stable angina pectoris.Methods Five hundred and seventy three patients underwent elective coronary angiography in a bi-racial cohort study,which included 295 patients with stable angina peetoris(SAP)and 278 subjects served as control.Plasma level of MPO and traditional risk factors of coronary artery disease(CAD)were measured.Results MPO levels did not differ significantly between control group and SAP group[126.3(95.8-160.2)mg/L vs 123.6(97.4-150.0) mg/L P＞0.05].MPO levels were similar across ethnicity and gender[black male 119.6(94.8-146.9) mg/L,white male 124.6(99.9～154.6)mg/L,black female 124.0(93.3～152.3)mg/L and white female 127.5(95.3～159.8)mg/L],and were correlated positively with the levels ofⅦfactor(r= 0.251,P＜0.01),fasting plasma glucose(r=0.095,P＜0.05),triglyceride(r=0.186,P＜0.01), total cholesterol(r=0.081,P＜0.05),high-sensitivity C-reactive protein(r=0.123,P＜0.01) and fibrinogen levels(r=0.077,P＜0.01),negatively correlated with adiponectin level(r=-0.115, P＜0.01).Conclusions Plasma MPO level is not elevated in patients with SAP.This suggests that MPO is not a characteristic feature of SAP.There are also no significant relationships between different genders and between different ethnicities.

Objective To study the clinical and imaging features of patients with bone metastases from breast cancer and identify the factors related to the incidence of bone metastases. Methods Three hundred and thirty-four patients with breast cancer were recruited into this study. Whole-body 99Tcm-methylene disphosphonate (MDP) bone scan, clinical staging, pathological, immunohistochemical and serological test results were analyzed retrospectively. χ2 test was used for statistical analysis. Results The incidence rate of bone metastases for patients with and without lymph node metastases was 71% (152/214) and 22. 5% (27/120), respectively (χ2 =72.80, P =0.000). The incidence rate of bone metastases from infiltrated non-specified and specified breast cancer was 69% (203/294) and 41.7% (5/12), respectively (χ2 =3. 97, P=0.046). Alkaline phosphatase (ALP) was elevated in 28.5% (51/179) and 14.9%(11/74) of patients with and without bone metastases, respectively (χ2 = 5. 25, P = 0.022 ). Carcinoembryonic antigen (CEA), carbohydrate antigen (CA) 15-3, CA125, CA19-9 increased in 68.7% ( 123/179) and 27.0% (20/74) of patients with and without bone metastases, respectively (χ2 = 37. 03, P =0. 000). Conclusions The incidence of bone metastases from breast cancer is correlated to pathological types of primary tumor and lymph node metastases. Bone metastases occurs more frequently in patients with infiltrated, non-specified, primary cancer and with lymph node metastases. Serum ALP, CEA, CA15-3,CA125, CA19-9 might be the tumor makers for early diagnosis of bone metastases from breast cancer.

Objective:To explore the value of echocardiography for diagnosing infectious endocarditis (IE).Methods:A total of 487 patients with cardiovascular implantable electronic devices (CIED) infection treated in our hospital from 2013-01 to 2015-06 were enrolled.Based on symptoms,blood culture and echocardiography,9 patients with suspected IE were further examined by 18F-FDG PET-CT to confirm their diagnosis and classification.Definitive therapy was conducted and the patients were followed-up for 1 year to confirm the diagnostic accuracy of echocardiography on CIED induced IE.Results:3 patients were preliminarily diagnosed for bacteremia since no vegetation was found by echocardiography,while IE was finally diagnosed by PET-CT.2 patients were preliminarily diagnosed for IE by echocardiography presented valvular vegetation,while PET-CT showed no evidence of vegetation;then one of them was diagnosed as bacteremia by positive blood culture and another was diagnosed as non-infection.4 patients were preliminarily diagnosed for IE by echocardiography indicated existing vegetation after CIED lead extraction,while PET-CT demonstrated no infection sign in heart chamber and the finally diagnosed was as "non-infectious fibrous residual tissue".According to final diagnosis,definitive therapies were performed to specific patients with at least 1 year follow-up study,no one had new and recurrent infection.Conclusion:Echocardiography had deficiency for diagnosing vegetation in heart chamber especially in suspicious IE patients after CIED lead extraction.It is necessary to make accurate diagnosis with other method for guiding appropriate therapy.

OBJECTIVETo investigate the relationship between complex 1311 mutation of C-->T in exon 11 and 93 T-->C in intron 11 of G6PD gene and the G6PD deficiency.

METHODSUsing NBT paper strip method to screen and quantitative NBT method to confirm G6PD deficiency. PCR-SSCP technique was used to find the abnormal exon 11 and the amplification refractory mutation system (ARMS) to identify 1311 mutation, and DNA sequencing to identify the complex mutation at 1311 in exon 11 and 93 in intron 11.

RESULTSAbnormal band in exon 11 was found in 12 cases. DNA sequencing showed that they were 1311 mutation together with 93 mutation.

CONCLUSIONThis complex mutation may be the cause of reduced activity of G6PD enzyme.

Base Sequence ; DNA Mutational Analysis ; Exons ; genetics ; Genetic Testing ; Glucosephosphate Dehydrogenase ; genetics ; Glucosephosphate Dehydrogenase Deficiency ; diagnosis ; enzymology ; genetics ; Humans ; Introns ; genetics ; Molecular Sequence Data ; Point Mutation ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide ; Polymorphism, Single-Stranded Conformational

OBJECTIVETo study the safety, biodistribution, and dosimetry of myocardial perfusion imaging agent 99Tc(m)N-NOET in 10 healthy volunteers.

METHODS744-792 MBq of 99Tc(m)N-NOET was injected to each volunteer. Safety parameters and adverse event was measured in 24 hours of injection. Biodistribution was studied by whole-body imaging 1, 30 minutes, 1, 2, 4, 8, and 24 hours after the injection of 99Tc(m)N-NOET. The estimation of dosimetry was based on the standard medical internal radiation dose method using MIRDOSE 3.0 analysis program. Myocardial single photon emission computed tomography (SPECT) imaging was performed at 1 and 4 hours after injection.

RESULTSNo undesirable effects were reported by the subject during 24 hours after injection of 99Tc(m)N-NOET. No clinically significant changes were found in vital signs (heart rate, blood pressure, and electrocardiogram). No biochemical aspects and serology changes were measured. The myocardial SPECT imaging was clear. Cardiac uptake of 99Tc(m)N-NOET was as high as 2.68% at 2 hours after injection. The heart to lung ratio was more than 1 from 30 minutes after injection, reaching a maximum of 1.91 +/- 0.53 at 2 hours after injection. Radiation dosimetry calculations indicated an effective absorbed dose of 1.28 x 10(-5) Sv/MBq. The dosimetry in each main organ is lower then 50 mGy given 740 MBq of 99Tc(m)N-NOET in once imaging.

CONCLUSIONS99Tc(m)N-NOET exhibits high cardiac uptake and low estimated effective absorbed dose. It's a safe myocardial perfusion imaging agent.

Heart ; diagnostic imaging ; Humans ; Myocardium ; metabolism ; Organotechnetium Compounds ; adverse effects ; pharmacokinetics ; Radiation Dosage ; Radiopharmaceuticals ; adverse effects ; pharmacokinetics ; Thiocarbamates ; adverse effects ; pharmacokinetics ; Tissue Distribution ; Tomography, Emission-Computed, Single-Photon

AIMTo investigate the regulatory function of Cox7a2 on steroidogenesis and the mechanism involved in TM3 mouse Leydig cells.

METHODSThe cDNA of Cox7a2 was cloned from TM3 mouse Leydig cells. It was subcloned to pDsRed-Express-N1 and transfected back into TM3 mouse Leydig cells for Cox7a2 overexpression by transient gene transfection. Steroidogenesis affected by overexpressed Cox7a2 was studied by ELISA. To elicit the mechanism of this effect, expression of steroidogenic acute regulatory (StAR) protein and reactive oxygen species (ROS) were examined by Western blot and fluorometer, respectively.

RESULTSThe cDNA of Cox7a2 (249 bp) was cloned from Leydig cells and confirmed by DNA sequencing. After constructed pDsRed-Express-N1-Cox7a2 was transfected back into TM3 mouse Leydig cells, Cox7a2 inhibited not only luteinizing hormone (LH)-induced secretion of testosterone but also the expression of StAR protein. At the same time, Cox7a2 increased the activity of ROS in TM3 mouse Leydig cells.

CONCLUSIONCox7a2 inhibited LH-induced StAR protein expression, and consequent testosterone production, at least in part, by increasing ROS activity in TM3 mouse Leydig cells.

Animals ; Cell Line ; Cloning, Molecular ; Electron Transport Complex IV ; genetics ; metabolism ; Leydig Cells ; drug effects ; metabolism ; Luteinizing Hormone ; pharmacology ; Male ; Mice ; Phosphoproteins ; metabolism ; Plasmids ; Reactive Oxygen Species ; metabolism ; Recombinant Proteins ; metabolism ; Testosterone ; metabolism ; Transfection

OBJECTIVETo study the expression and localization of ATP50 by construction of ATP50-pEYFP-N1 in primary cultured mouse Leydig cells.

METHODSPrimary cultured mouse Leydig cells were confirmed by 3B-HSD staining. ATP50 was cloned into pEYFP-N1 between Bam HI and Eco RI sites. Cell-transfection and living-cell fluorescence imaging microscopy were employed to investigate the sub-cellular localization of YFP-ATP50 in TM3 mouse Leydig cells.

RESULTSATP50 green fluorescent protein was well co-localized with red fluorescence mitochondrion marker-Mitotracker in TM3 mouse Leydig cells.

CONCLUSIONATP50 was expressed in primary cultured mouse Leydig cells. The fluorescent expression vector of ATP50 was constructed successfully and YFP-ATP50 was located in mitochondria in TM3 mouse Leydig cells, which provided a useful clue for further research on the steroidogenesis dysfunction in aging males.

Adenosine Triphosphatases ; biosynthesis ; genetics ; Animals ; Carrier Proteins ; biosynthesis ; genetics ; Cells, Cultured ; Cloning, Molecular ; Genetic Vectors ; Green Fluorescent Proteins ; biosynthesis ; genetics ; Leydig Cells ; metabolism ; Male ; Membrane Proteins ; biosynthesis ; genetics ; Mice ; Mitochondria ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; Transfection

ObjectiveTo observe the morphological changes of bone in the progress of chronic fluorosis.MethodsWistar rats were randomly divided into three groups,30 rats in each group:normal control group,experimental group Ⅰ and experimental group Ⅱ according to body weight.Rats in normal control group drank distilled water freely.Experimental group Ⅰ and group Ⅱ drunk distilled water with sodium fluoride preparation of fluorine containing ion 100,150 mg/L solution for six months,respectively.Bone mineral density was detected by X-ray,bone morphological changes were observed under light microscope and bone histomorphometric parameters were calculated using image analysis software.ResultsThe bone mineral density values were different statistically between the three groups after feeding for 2 and 4 months(F =19.79,3.28,all P ＜ 0.05).However no significant difference was found after feeding for 6 months(F =1.80,P ＞ 0.05).The bone mineral density of experimental group Ⅰ (0.20 ± 0.03,0.21 ± 0.03) was significantly higher than that of the normal control group(0.17 ± 0.03,0.20 ± 0.04) after feeding for 2 and 4 months.The bone mineral density of experimental group Ⅱ (0.21 ± 0.02) was lower than that of normal control group(0.22 ± 0.03) after feeding for 6 months.The bone lamella in experimental group Ⅰ was arranged disorderly,the number of osteocytes increased with their nucleus atrophy and the osteoblasts were more than that of control grouo which arranged in layers observed under light microscooy.In exoerimental group Ⅱ,the bone lamella was bent deformation,the number of osteocytes had decreased with their nucleus shrinking or even disappeared and the number of osteoclasts had increased significantly observed under light microscopy.In experimental group Ⅰ,the mean trabecular density [(0.33 ± 0.03)％] increased and the mean trabecular separation,thickness [( 163.57 ± 1.99),(59.26 ± 7.18 ) μm] decreased compared with that of normal control group [(0.31 ± 0.02)％,(186.60 ± 2.90)μm,(86.42 ± 1.48)μm,all P ＜ 0.05].In experimental group Ⅱ,the mean trabecular density[(0.26 ± 0.02)％] decreased,the mean trabecular thickness[(71.42 ± 10.77)μm] reduced compared with that of normal control group[(0.31 ± 0.02)％,(86.42 ± 1.48)μm].ConclusionsExcess fluoride can damage bone tissue.Low doses of fluoride can stimulate osteoblast activity and enhance osteogenesis.The activity of osteoblasts is great than that of osteoclasts.High doses of fluoride can stimulate both osteoblasts and osteoclasts activity,but mainly the activity of osteoclasts,and bone resorption increases.