BACKGROUND: The study aimed to explore the effects of hypothermia state induced by 4 oC normal saline (NS) on liver biochemistry, enzymology and morphology after restoration of spontaneous circulation (ROSC) by cardiopulmonary resuscitation (CPR) in swine. METHODS: After 4 minutes of ventricular fibrillation (VF), standard CPR was carried out. Then the survivors were divided into two groups: low temperature group and normal temperature group. The low temperature (LT) group (n=5) received continuously 4 oC NS at the speed of 1.33 mL/kg per minute for 22 minutes, then at the speed lowering to 10 mL/kg per hour. The normal temperature (NT) group (n=5) received NS with normal room temperature at the same speed of the LT group. Hemodynamic status and oxygen metabolism were monitored and the levels of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) were measured in blood samples obtained at baseline and at 10 minutes, 2 hours and 4 hours after ROSC. At 24 hours after ROSC, the animals were killed and the liver was removed to determine the Na+-K+-ATPase and Ca2+-ATPase enzyme activities and histological changes under a light or electron microscope. RESULTS: Core temperature was decreased in the LT group (P<0.05), while HR, MAP and CPP were not significantly decreased (P>0.05) compared with the NT group (P>0.05). The oxygen extraction ratio was lower in the LT group than in the NT group (P<0.05). The serum levels of ALT, AST and LDH increased in both groups but not significantly in the LT group. The enzyme activity of liver ATP was much higher in the LT group (Na+-K+-ATP enzyme: 8.64±3.32 U vs. 3.28±0.71 U; Ca2+-ATP enzyme: 10.92±2.12 U vs. 2.75±0.78 U, P<0.05). The LT group showed less cellular edema, inflammation and few damaged mitochondria as compared with the NT group. CONCLUSION: These data suggested that infusing 4 oC NS continuously after ROSC could quickly lower the core body temperature, while maintaining a stable hemodynamic state and balancing oxygen metabolism, which protect the liver in terms of biochemistry, enzymology and histology after CPR.

BACKGROUND: The association of genetic variation in the IRAK-1 gene with sepsis outcome has been proved. However, few studies have addressed the impact of the IRAK-4 gene variants on sepsis risk. This study aimed to determine whether the polymorphisms in the IRAK-4 gene are associated with susceptibility to and prognosis of severe sepsis in the Chinese Han ethnic population.METHODS: In this case-control study, 192 patients with severe sepsis hospitalized in the emergency department of Zhongshan Hospital from February 2006 to December 2009 and 192 healthy volunteers were enrolled. Exclusion criteria included metastatic tumors, autoimmune diseases, AIDS or treatment with immunosuppressive drugs. This study was approved by the ethical committee of Zhongshan Hospital, Fudan University. Sepsis patients were divided into a survival group (n=124) and a non-survival group (n=68) according to the 30-day mortality. Primer 3 software was used to design PCR and sequencing primers. Genomic DNA was extracted from peripheral blood mononuclear cells. Seven tagSNPs in IRAK-4 were selected according to the data of the Chinese Han population in Beijing from the Hapmap project and genotyped by direct sequencing. The chi-square test was used to evaluate the differences in genotype and allele frequencies between the two groups.RESULTS: The distributions of all tagSNPs were consistent with Hardy-Weinberg equilibrium. The allele and genotype frequencies of rs4251545 (G/A) were significantly different between the severe sepsis and healthy control groups (P=0.015, P=0.035, respectively). Carriers of the rs4251545A had a higher risk for severe sepsis compared with carriers of the rs4251545G (OR=1.69, 95％ CI: 1.10-2.58). The allele and genotype frequencies of all SNPs were not significantly different between the survival group and non-survival group.CONCLUSION: These findings indicate that the variants in IRAK-4 are significantly associated with susceptibility to severe sepsis in the Chinese Han ethnic population.

Objective To investigate the brain mechanisms of aging of the visual attention. Methods Through the precue-target visual search paradigm using the event-related potentials technique,16 young and 16 elderly subjects participated the electrophysiological experiment.The background was comprised of three homocentric black circles and eight English capital letters consisted of each circle.The letter “T” was designed as the target stimulus.T was a target only when it located the circle clued to the same size of the precue.For example,when the cue was the “large”,“T” may appear within 3 circles.When the cue was the “median”,“T” may appear within either the median or small circles.When the cue was small,the target “T” may appear only within the small circle. Results The reaction time of the two groups of subjects became quick with the reduction of the cue scale,while the amplitudes of P1 and N1 components of event-related potentials increased with the decrease of the cue scale.Old subjects showed longer response time than did young subjects,and the posterior P1 component was enhanced significantly and N1 component was inhibited obviously.The P2 component was manifested as significantly inhibitory effect not only in the amplitude but also in the abnormal and unstable waveform.Conclusions The cognitive function of elderly subjects declines in the research of target stimuli,which suggests that the age-related changes could lead to deficit in the posterior area of the brain to visual spatial attention (involuntary attention).

OBJECTIVETo obtain massive human pancreatic islets with modified techniques and evaluation of the islets for the clinical allo-transplantation to treat type I and II diabetes.

METHODS28 consecutive adult human pancreata were isolated with modified automated techniques. Islets were purified using continuous density gradient. The islet yield was counted with international standard known as islet equivalent (IEQ). The function of the isolated islets was evaluated by measuring DNA/insulin ratio, static glucose stimulating test in vitro and transplanting the islets into diabetic nude mice in vivo followed by abdominal glucose tolerance test and C peptide measurement.

RESULTSThe yield of 28 consecutive human pancreata isolations ranged from 5 000 to 1 030 000 IEQs/pancreas with the average of 291 635 IEQs/pancreas. The first 13 isolations yielded 49 123 IEQs/pancreas, 846 IEQs/g and, purity 87% in average. The remained 15 isolations after the modifications yielded 501 813 IEQs/pancreas, 7 003 IEQs/g and purity 89% in average. The results of in vitro SGS showed good response to the different glucose concentration. 34 diabetic nude mice were transplanted under the renal capsule with the freshly isolated islets. 29 out of 34 diabetic mice obtained normoglycemia within 12 hours and the glucose tolerance tests were near normal. Serum C peptide level of transplanted mice is close to that of the control group.

CONCLUSIONSMassive human islets can be isolated with the modified techniques. Quality assessment of these islets both in vitro and in vivo has indicated that these high quality human islets could be used for the clinical allogeneic islet transplantation.

Adult ; Animals ; Cell Separation ; methods ; Diabetes Mellitus, Experimental ; surgery ; Glucose ; Humans ; In Vitro Techniques ; Islets of Langerhans ; cytology ; drug effects ; physiology ; Islets of Langerhans Transplantation ; Mice ; Mice, Nude ; Transplantation, Heterologous

Objective To examine the mutagenicity and antioxidant activity of Proanthocyanidin( PC). Methods Three mutagenicity tests including Ames test,bone marrow micronucleus test and sperm malformation test in mice were conducted. According to the level of erythrocyte MDA,11-month-old rats were randomly divided into five groups including blank control,solvent control and 41. 67,83. 33,250. 0 mg/kg test groups. The content of erythrocyte MDA,SOD and GSH-PX activity of each dosage group were then observed after administration of PC to test the antioxidant activity. Results In all three mutagenicity tests,no mutagenic effect was observed in any PC - treated group. Compared with two control groups,the content of erythrocyte MDA was significantly decreased in 83. 33 and 250. 0 mg/kg groups(both P<0. 05) and GSH-Px activity was significantly increased in 250 mg/kg group(P<0. 05). Conclusion PC had no mutagenic effect but showed antioxidant activity under our experimental conditions.

Objective To explore the mutagenic effect of sodium pentachlorophenate (NaPCP) on zebrafish p53 gene coding sequence(CDS) in somatic cell.Methods The experiment was carried out using tuebingen strain of zebrafish, according to the results of acute toxicity test to determine the exposure levels in zebrafish.Zebrafish were randomly divided into blank control group and exposed groups, each containing 10 zebrafish.After exposing for 45d of NaPCP, the RNA was extracted from liver of zebra fish, and the p53 gene including a complete coding sequence of was obtained by RT-PCR.Results LC50 of NaPCP was 18.4 μg/L.Sequence analysis showed that the p53 gene CDS length of 1125bp, encoding 374 amino acids.The percent identity between the published zebrafish sequence of p53 (GI:425876786)and ours was 99.2%,with the other biological sequence of p53 existing some differences.After 45d exposure, zebrafish p53 gene of NaPCP exposure group had mutated at the concentration of 1.8 μg /L.The base substitution of GAG→AAG at codon 8,CAT→CAG at codon 148 and CAG→CAA at codon 229 were detected by PCR-directed sequencing.This may result in the Glu→Lys and His→Gln of expressed p53 protein.Conclusion NaPCP is a kind of gene mutation, which can induce the mutation of p53 gene in zebrafish somatic cells, that has the potential mutagenic risk for humans.

Objective To learn the changes of blood lead levels and serum biochemical parameters of the school agechildren from different areas. Methods All research objects, the school age children, were from three different areasrespectively, including a mountainous area (L area), an island area (H area) where there is not history of Pb pollution,and an industry area (N area) in relation to Pb pollution. The morning urine and peripheral venous blood samples werecollected from the school age children. Pb in blood (PbB), δ-aminoaevulinic acid in urine (ALA), Ca2+, BUN, Cr inserum, and Thyroid Stimulating Hormone (TSH), thyroxin (T4), free thyroxin (FT4) levels were detected. ResultsPbB levels [M was 36.0 ppb] of the school age children from N area were significantly higher than that of L area [22.0 ppb] andH area [23.8 ppb]. On the contrary, serum Ca2+ levels of the school age children from N area were significantly lower than thatof L area and H area. Serum T4 of N area was significantly lower than that of L area and H area. Serum FT4 of H area wassignificantly higher than that of L area and N area. And TSH of N area and H area were both obviously lower than that of L area.But all of these thyroxin indexes were in the range of normal values. Conclusion It should be widely concerned that thesignificant elevation of PbB levels may have a negative impact on school age children in the future.

BACKGROUNDSterol regulatory element binding protein (SREBP)-2 plays a key role in lipid homeostasis by stimulating gene expression of cholesterol biosynthetic pathways. The insulin-like growth factor binding protein (IGFBP) family regulates growth and metabolism, especially bone cell metabolism, and correlates with osteonecrosis. However, association of their gene polymorphisms with risk of avascular necrosis of the femoral head (ANFH) has rarely been reported. We determined whether SREBP-2 and IGFBP-3 gene polymorphisms were associated with increased ANFH risk in the Chinese population.

METHODSTwo single nucleotide polymorphisms of SREBP2 gene, rs2267439 and rs2267443, and one of IGFBP-3 gene, rs2453839, were selected and genotyped in 49 ANFH patients and 42 control individuals by direct sequencing assay.

RESULTSThe frequencies of rs2267439 TT and rs2267443 GA of SREBP2 and rs2453839 TT and CT of IGFBP-3 in the ANFH group showed increased and decreased tendencies (against normal control group), respectively. Interaction analysis of genes revealed that the frequency of carrying rs2267439 TT and rs2267443 GA genotypes of SREBF-2 in ANFH patients was significantly higher than in the control group (P < 0.05). Association analysis between polymorphisms and clinical phenotype demonstrated that the disease course in ANFH patients with the rs2453839 TT genotype of IGFBP-3 was significantly shorter than that of CT + CC carriers (P < 0.01). CT + CC genotype frequency in patients with stage III/IV bilateral hip lesions was significantly higher than in those with stage III/IV unilateral lesions and stage II/III bilateral lesions (P < 0.05 - 0.02).

CONCLUSIONSOur results suggested that interaction of SREBP-2 gene polymorphisms and the relationship between the polymorphisms and clinical phenotype of IGFBP-3 were closely related to increased ANFH risk in the Chinese population. The most significant finding was that the CT + CC genotype carriers of IGFBP-3 rs2453839 were highly associated with the development of ANFH.

Adult ; Asian Continental Ancestry Group ; genetics ; Female ; Femur Head Necrosis ; genetics ; Genetic Predisposition to Disease ; Humans ; Insulin-Like Growth Factor Binding Protein 3 ; genetics ; Male ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Genetic ; genetics ; Sterol Regulatory Element Binding Protein 2 ; genetics

OBJECTIVETo investigate intake of water in different periods of a day of primary and middle school students in Beijing, Shanghai, Guangzhou and Chengdu of China.

METHODSA total of 5914 primary and middle school students from Beijing, Shanghai, Guangzhou and Chengdu were recruited using multiple-stage random sampling method from September to October 2011. Among them, 5868 students completed the survey. The information of amounts and types of drinking in 8 time periods, morning, afternoon, night as well as in meal and non-meal time in 24 hours of a day was recorded by subjects for seven consecutive days using a quantitative measurement. This information was analyzed by different periods.

RESULTSThe amount of drinking water among 8 periods of a day was (99 ± 101), (130 ± 106), (191 ± 155), (96 ± 107), (246 ± 172), (90 ± 101), (188 ± 135) and (50 ± 81) ml, respectively for before breakfast, during breakfast, after breakfast, during lunch, after lunch, during dinner, after dinner, and midnight (F = 1679.77, P < 0.01). Based on three periods of a day, the amount of drinking water in the morning (420 ± 242) ml was the statistically significantly most, followed by afternoon (341 ± 199) ml, and the least in the evening (327 ± 195) ml (F = 325.23, P < 0.01). The distribution trend of plain water in three periods was the same as total drinking water, with (270 ± 209), (250 ± 179) and (224 ± 177) ml, respectively (F = 84.89, P < 0.01) ; but beverage consumption was the most in the morning(151 ± 133) ml, then in the evening (103 ± 122) ml, and the least in the afternoon (91 ± 199) ml (F = 373.56, P < 0.01). The daily plain water consumption in meal time was (316 ± 247) ml, while it in non-meal time was (773 ± 445) ml with statistically significant difference (Z = 65.2, P < 0.01). There was no significant difference in meal time between plain water (165 ± 194) ml and beverage (151 ± 152) ml (Z = 0.59, P = 0.56) whilst plain water (579 ± 408) ml in non-meal time was significantly higher than beverages (194 ± 204) ml (Z = 64.5, P < 0.01).

CONCLUSIONThe amount of drinking water of primary and middle school students in different periods of a day was different in four cities of China.

Adolescent ; Beverages ; Child ; China ; Diet Surveys ; Drinking Behavior ; Drinking Water ; Female ; Humans ; Male ; Students ; Surveys and Questionnaires ; Time Factors ; Urban Population

BACKGROUNDSeveral isoforms of p53 have been reported, which may have varying functions and expressions. This study aimed to analyze the expression patterns of p53 isoforms in renal cell carcinoma (RCC) at the mRNA and protein levels and their associations with clinical and pathologic factors to explore the mechanism of p53 isoforms' activity in RCC.

METHODSThe specimens of tumours (T) and clinically normal tissues (N) adjacent to them were collected from 41 patients with RCC. mRNA expression levels of p53 isoforms were detected using RT-PCR followed by nested PCR. Protein expression levels were detected using immunohistochemisty and Western blotting with the anti-p53 antibodies DO-1 and DO-12. The data were analyzed with clinicopathological features by chi(2) test or Fisher's exact test.

RESULTSp53 mRNA was expressed in all tumours and matched clinically normal tissue adjacent to the tumour. All six isoforms could be detected in tumour and normal tissues, with the exception of the Delta133p53beta isoform, which was not detected in the normal tissue. Of the six isoforms, p53beta mRNA was significantly overexpressed in tumour samples (P < 0.001), and correlated with tumour stage. Nested PCR results consistently indicated the presence of p53gamma (19T/22N), Delta133p53 (33T/26N), Delta133p53beta (2T/0N), and Delta133p53gamma (13T/9N). Immunohistochemical analysis showed that p53 was expressed only in tumour tissues and correlated with tumour stage and grade. The results of Western blotting analysis were consistent with these findings.

CONCLUSIONSAlthough all six isoforms are present in RCC, their function in tumour development or progression might be different. Our findings suggest that p53beta might play an important role in the formation of RCC and it might be used as a new predictor and therapeutic target for RCC.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Blotting, Western ; Carcinoma, Renal Cell ; genetics ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; genetics ; physiology ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; Protein Isoforms ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Suppressor Protein p53 ; genetics ; metabolism ; Young Adult