Objective To investigate the self-assembly and cellular localization patterns of fila-mentous temperature-sensitive protein Z (FtsZ) in Escherichia coli (E.coli) strains by using FtsZP74R, FtsZG77D and FtsZA81R mutants.Methods YFP or His labeled FtsZ proteins and the plasmids of FtsZ mu-tants were constructed by using molecular clone and site-directed mutagenesis methods.The targeted proteins were purified by affinity chromatography.FL37(△ftsZ-Cat) strains were constructed via linear DNA homol-ogous recombination.Living cell imaging was performed to observe the cellular localization patterns of FtsZ protein and its mutants in E.coli strains.The interactions between FtsZ-FtsZ/FtsZ mutants were examined by coi-mmunoprecipitation assay . The polymerization properties of FtsZ mutants were analyzed by light scattering.The activities of GTPase were monitored by using high performance liquid chromatography.Re-sults The P74, G77 and A81 amino acids were respectively replaced by different polar amino acids to change the amphipathicity of the helix within the domain of FtsZ ( 74-82 ) .The YFP-labeled FtsZP74R , FtsZG77D and FtsZA81R mutants failed to assemble into functional Z-ring structure in E.coli strains.The inter-actions between FtsZ protein and its mutants were weakened or completely disappeared.In addition, in vitro experiments showed that P74R, G77D and A81R mutations caused a decrease in the polymerization efficien-cy of FtsZ monomer.The activity of GTPase was significantly decreased in the FtsZA81R mutant. Conclusion The P74, G77 and A81 were critical amino acids in the function and assembly of FtsZ protein in E.coli strains.Moreover, A81 amino acid regulated the lateral interaction of FtsZ monomer and the activity of GTPase.

Objective:To investigate the recognition and injury prevention strategies of hepatic artery variations during hepatic portal lymphadenectomy. Methods:A retrospective analysis was performed, and 12 patients of hepatic arterial variation among 62 pa-tients with hepatic portal lymphadenectomy were the subjects. The study was conducted in the First Affiliated Hospital of Bengbu Medi-cal College between January 2013 and July 2014. The intraoperative treatment and postoperative complications were recorded. Results:Among 12 cases of hepatic artery variation, we found the following cases:3 cases (25.0%) of Michels' Type III, 2 cases (16.7%) of Mi-chels' Type VI, 1 case (8.3%) of Michels' Type IX, 1 case (8.3%) of Hiatt's Type 6, 2 cases (16.7%) of spatial location variation between right hepatic artery and hepatic duct, 2 cases (16.7%) of left and right hepatic artery originating from a common hepatic artery, and 1 case (8.3%) of right hepatic artery originating from the gastroduodenal artery. No injury of hepatic artery occurred. Two cases had post-operative complications, including 1 case of pancreatic leakage and 1 case of incision infection;postoperative hemorrhage, bile leakage, hepatic abscess did not occur in these two cases. Patients recovered well in general. Conclusion:Hepatic arterial injury can be signifi-cantly reduced by the following:increased familiarity with the various types of hepatic artery variations;complete imaging examina-tions for inspection and evaluation before surgery;and careful and meticulous operations in surgery.

Adenocarcinoma ; metabolism ; pathology ; surgery ; Adenocarcinoma, Clear Cell ; metabolism ; pathology ; Adult ; CA-125 Antigen ; metabolism ; Carcinoembryonic Antigen ; metabolism ; Cervical Intraepithelial Neoplasia ; metabolism ; pathology ; surgery ; Cystadenocarcinoma, Serous ; metabolism ; pathology ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Humans ; Hysterectomy ; Ki-67 Antigen ; metabolism ; Lymph Node Excision ; Membrane Proteins ; metabolism ; Neoplasm Invasiveness ; Uterine Cervical Neoplasms ; metabolism ; pathology ; surgery

OBJECTIVETo investigate the relationship between the second to the fourth digit ratio (2D:4D) and body mass index (BMI) in infertile men of the Han ethnic group in Ningxia.

METHODSUsing anthropometry, we calculated the mean ratio of 2D:4D and BMI of 197 infertile men and 148 normal healthy male controls, followed by analysis of their relationship.

RESULTSThe BMI was correlated positively with the 2D:4D ratio of the left hand in the infertile men (P < 0.05) and in the patients with a higher 2D:4D ratio of the left hand (P < 0.05), but negatively with the 2D:4D ratio of the righ/left (Dr-1) (left: P < 0.01; Dr-l: P < 0.05). The mean 2D: 4D ratio and BMI were both lower in the normal control than in the infertile men, with statistically significant differences in BMI (P < 0.05) and the 2D:4D ratio of the left hand (P < 0.05).

CONCLUSIONThere is a correlation between the 2D:4D ratio and BMI in infertile men.

Body Mass Index ; Case-Control Studies ; Fingers ; anatomy & histology ; Humans ; Infertility, Male ; diagnosis ; Male

OBJECTIVETo investigate the dynamic variation and action mechanism of sICAM-1 and p38 mitogen-activated protein kinases (MAPK) signal transduction in human severe trauma and resuscitation, as well as the effect of lactated Ringer's solution(LR), 7.5% sodium chloride solution(HS) and 20% albumin injection(ALB) on the incidence and mortality of multiple organ dysfunction syndrome (MODS).

METHODSSeventy-two severe trauma patients (ISS score 16-43) were divided into ISS < or = 25 and ISS > 25 groups (each group was subdivided into LR, HS and ALB groups). ELISA was used to measure the concentration of sICAM-1. Western blot was used to measure the expression of p38 MAPK.

RESULTSCompared with LR group, the transfusion volume needed for maintaining systolic blood pressure > or = 90 mm Hg was significantly decreased in HS and ALB groups (P < 0.05). Compared with the control group, the concentration of blood sICAM-1 and the expression of p38 MAPK was elevated from 4 to 48 hours after trauma in all experimental groups (P < 0.05-0.01). At 4, 12, and 24 hours, there was significant correlation between the expression of p38 MAPK and sICAM-1 (P < 0.01). Compared with LR group, sICAM-1 and p38 MAPK in HS and ALB groups were decreased (P < 0.05). sICAM-1 and p38 MAPK were significantly higher in the group of ISS > 25 than that of ISS < or = 25 (P < 0.05). MODS incidence and mortality were significantly higher in the group of ISS > 25 than that of ISS < or = 25 (P < 0.05). MODS incidence and mortality were lower in HS and ALB groups than LR group (P < 0.05).

CONCLUSIONSThe up-regulation of polymorphonuclear neutrophil-endotheliocytes (PMN-EC) adhesion may be due to the increased sICAM-1 expression during severe trauma. The up-regulation of sICAM-1 expression is correlated with the activation of p38 MAPK. During severe trauma, the levels of sICAM-1 and p38 MAPK, as well as the incidence and mortality of MODS are lower when HS and ALB are used than single lactated LR solution is used.

Adult ; Albumins ; therapeutic use ; Cell Adhesion ; Female ; Fluid Therapy ; methods ; Humans ; Intercellular Adhesion Molecule-1 ; blood ; physiology ; Isotonic Solutions ; therapeutic use ; Male ; Middle Aged ; Multiple Organ Failure ; epidemiology ; mortality ; Resuscitation ; Saline Solution, Hypertonic ; therapeutic use ; Systole ; Wounds and Injuries ; blood ; physiopathology ; therapy ; p38 Mitogen-Activated Protein Kinases ; blood ; physiology

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

BACKGROUNDIn prokaryotic organisms, the mechanism responsible for the accurate partition of newly replicated chromosomes into daughter cells is incompletely understood. Segregation of the replication terminus of the circular prokaryotic chromosome poses special problems that have not previously been addressed. The aim of this study was to investigate the roles of several protein components (MreB, MreC, and MreD) of the prokaryotic cytoskeleton for the faithful transmission of the chromosomal terminus into daughter cells.

METHODSStrain LQ1 (mreB::cat), LQ2 (mreC::cat), and LQ3 (mreD::cat) were constructed using the Red recombination system. LQ11/pLAU53, LQ12/pLAU53, LQ13/pLAU53, LQ14/pLAU53, and LQ15/pLAU53 strains were generated by P1transduction of (tetO) 240 -Gm and (lacO) 240 -Km cassettes from strains IL2 and IL29. Fluorescence microscopy was performed to observe localization pattern of fluorescently-labeled origin and terminus foci in wild-type and mutant cells. SOS induction was monitored as gfp fluorescence from PsulA-gfp in log phase cells grown in Luria-Bertani medium at 37°C by measurement of emission at 525 nm with excitation at 470 nm in a microplate fluorescence reader.

RESULTSMutational deletion of the mreB, mreC, or mreD genes was associated with selective loss of the terminus region in approximately 40% of the cells within growing cultures. This was accompanied by significant induction of the SOS DNA damage response, suggesting that deletion of terminus sequences may have occurred by chromosomal cleavage, presumably caused by ingrowth of the division septum prior to segregation of the replicated terminal.

CONCLUSIONSThese results imply a role for the MreBCD cytoskeleton in the resolution of the final products of terminus replication and/or in the specific movement of newly replicated termini away from midcell prior to completion of septal ingrowth. This would identify a previously unrecognized stage in the overall process of chromosome segregation.

Chromosome Segregation ; genetics ; physiology ; Cytoskeleton ; metabolism ; Escherichia coli ; genetics ; metabolism

Since late 2010, porcine epidemic diarrhea virus (PEDV) has been re-emerging in central China. To explore the possible reason of the PEDV outbreaks, twelve PEDV field strains were isolated from different swine breeding farms in central China during 2010-2012, and molecular diversity, phylogenetic relationships of these strains with other PEDV reference strains were investigated. Sequence analysis of S, M and ORE3 genes revealed that the central China PEDV isolates had several specific nucleotides and amino acids which were different from PEDV reference strains. In addition, the entire S genes of eleven central China PEDV isolates were found to be nine nucleotides longer in length than CV777 and large number of amino acid variations was accumulated in the N-terminal region of S gene. Phylogenetic analysis showed that the central China PEDV isolates had close relationship with Korea strains (2007-2009), Thailand strains (2007-2008), Vietnam strains (2009-2010), Japan strains (2010), and other prevailing strains from other parts of China (2010-2012). However, they differed genetically from European strains (CV777, Brl/87), China strains (2003-2007) and the vaccine strains (CV777) used in China. These results imply that a rapid variation and evolution of central China PEDV strains has occurred in recent years, and a more efficient vaccine strain should be selected to prevent and control outbreaks of PEDV in China.
Animals ; China ; epidemiology ; Disease Outbreaks ; Feces ; virology ; Molecular Sequence Data ; Open Reading Frames ; Phylogeny ; Porcine epidemic diarrhea virus ; classification ; genetics ; isolation & purification ; Swine ; Swine Diseases ; epidemiology ; virology ; Viral Proteins ; genetics

OBJECTIVESTo explore intervention with electromagnetic noise for co-suppression effect on gap-junctional intercellular communication (GJIC) induced or strengthened by low intensity magnetic field with carcinogen 12-O-tetradecanoylphorbol-13-acetate (TPA).

METHODSFibroblast cells from NIH 3T3 mice were exposed to extremely low intensity magnetic field (MF) 0.2 mT, 0.2 mT + TPA or/and electromagnetic noise with the same intensity of MF for 24 h, and GJIC was determined using fluorescence recovery analysis after photobleaching (FRAP) with a laser-scanning confocal microscope (Leica, Germany).

RESULTSGJIC function could be co-suppressed by MF of 0.2 mT with TPA, with fluorescence recovery of (23 +/- 11)%, lower than that in the control group [(46 +/- 19)%] and in the group with TPA only [(34 +/- 17) %] (P < 0.01), indicating 0.2 mT MF plus TPA could co-inhibit GJIC (P < 0.01). Superposition of 0.2 mT noise MF could get a fluorescence recovery of (35 +/- 19)% and significantly antagonize its co-suppression by TPA.

CONCLUSIONElectromagnetic noise of 0.2 mT could block the intensifying effect of power frequency magnetic field on TPA-induced GJIC inhibition.

Animals ; Cell Communication ; drug effects ; physiology ; radiation effects ; Cell Line ; Electromagnetic Fields ; adverse effects ; Fluorescence Recovery After Photobleaching ; Gap Junctions ; drug effects ; physiology ; radiation effects ; Mice ; NIH 3T3 Cells ; Noise ; adverse effects ; Tetradecanoylphorbol Acetate ; pharmacology

BACKGROUNDIn China, patients with hepatitis C virus (HCV)-associated liver disease are getting older, and thus the number of deaths due to such disease is increasing. The efficacy of combination therapy with ribavirin and interferon for chronic HCV infection in elderly patients has not been fully clarified. The aim of the present study was to evaluate the efficacy and tolerability of the combination therapy in the elderly patients.

METHODSSixty-eight chronic hepatitis C patients, who received the combination therapy, were classified into two age groups: elderly group ((3)60 years, n = 25) and non-elderly group (< 60 years, n = 43). Rapid virological response, complete early virological response, sustained virological response, relapse, non-response rate, and safety were compared between the elderly group and non-elderly group.

RESULTSOverall sustained virological response was lower in the elderly group than non-elderly group (44% vs. 75%, P = 0.012, OR = 0.270, and 95%CI 0.095 - 0.768). Among patients with HCV genotype 1, sustained virological response was lower in the elderly group than non-elderly group (45% vs. 77%, P = 0.015, OR = 0.247, 95%CI 0.078 - 0.781). The proportions of dose reduction due to laboratory abnormalities were significantly higher in the elderly group than non-elderly group (60.0% vs. 32.6%, P = 0.027). Multiple binary Logistic regression analysis confirmed that patient age was an associated factor for sustained virological response.

CONCLUSIONAmong patients with HCV genotype 1, the elderly patients had lower sustained virological response than non-elderly patients during pegylated interferon-alpha-2a plus ribavirin combination therapy.

Adult ; Aged ; Antiviral Agents ; therapeutic use ; Female ; Hepatitis C, Chronic ; drug therapy ; Humans ; Interferon-alpha ; therapeutic use ; Logistic Models ; Male ; Middle Aged ; Polyethylene Glycols ; therapeutic use ; Recombinant Proteins ; therapeutic use ; Ribavirin ; therapeutic use ; Young Adult