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 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

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

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

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

AIMTo identify a novel alternative transcript of the novel retinal pigment epithelial cell gene (NORPEG) expressed in the human testis.

METHODSA human testis cDNA microarray was established and hybridized with cDNA probes from human fetal testes, adult testes and human spermatozoa. Differentially expressed clones were sequenced and analyzed. One of these clones was a short transcript of NORPEG which we proceeded to analyze by RT-PCR.

RESULTSThe novel short alternative transcript of NORPEG was isolated and named sNORPEG. It was 3486 bp in length and contained a 2952-bp open reading frame, encoding a 110.4-kDa protein of 983 amino acids. Amino acid sequence analysis showed that the sNORPEG protein contains six ankyrin repeats and two coiled-coil domains. It shares a high homology with the NORPEG and ankycorbin proteins in both its sequence and motifs. Blasting the human genome database localized sNORPEG to human chromosome 5p13.2-13.3. Expression profiles showed that sNORPEG was expressed in human fetal testes, adult testes and spermatozoa. Moreover, sNORPEG was found to be ubiquitously expressed in human tissues.

CONCLUSIONsNORPEG is expressed in different developmental stages of the testis and encodes a protein that may have roles in human testis development and spermatogenesis.

Alternative Splicing ; Amino Acid Sequence ; Base Sequence ; Cytoskeletal Proteins ; genetics ; DNA, Complementary ; Gene Expression Profiling ; Humans ; Male ; Molecular Sequence Data ; Open Reading Frames ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Homology, Amino Acid ; Testis ; metabolism ; Transcription Factors ; genetics

OBJECTIVETo determine whether patients infected with chronic hepatitis C (CHC) show a differential distribution profile of IL-28B polymorphisms according to the presence of concomitant cryoglobulinemia.

METHODSSixty-two consecutive CHC patients were enrolled in the study between December 2008 and December 2010. All patients received combination therapy of pegylated interferon alpha-2a (weekly, 180 g, subcutaneous injection) plus ribavirin (daily, 10to15 mg/kg body weight, oral) for 48 weeks, with individualized dosage adjustments according to the patient's clinical situation. Cryoglobulins were detected visibly by separation of cryoprecipitates in patient serum samples. Three IL-28B SNPs (rs8099917, rs12979860, and rs12980275) were detected by sequencing. Response to treatment was assessed by measuring serum levels of HCV RNA by quantitative PCR at baseline (prior to treatment initiation), during treatment (4 and 12 weeks after treatment initiation), end of therapy (48 weeks after treatment initiation), and post-treatment (24 weeks after end of therapy). The significance of between-group differences were assessed by the Chi-square and Fisher's exact tests.

RESULTSCryoglobulinemia was detected in 43.5% (27/62) of the CHC patients and showed a female bias (59.3% vs. males: 34.3%, P = 0.05). Compared to CHC patients without cryoglobulinemia, the CHC patients with cryoglobulinemia showed significantly higher levels of HCV RNA at baseline (5.64+/-1.20 vs. 6.37+/-0.67, P less than 0.05) but lower frequencies of the IL28B rs8099917 TT genotype (94.3% vs. 63.0%, P = 0.002), rs8099917 T allele (97.1% vs. 81.5%, P = 0.003), and rs12979860 C allele (94.3% vs. 83.3%, P = 0.048). CHC patients with cryoglobulinemia and having the rs8099917 TT, rs12979860 CC, or rs12980275 AA genotype achieved a higher rate of sustained virological response.

CONCLUSIONCryoglobulinemia in CHC patients is associated with a differential distribution of IL-28B polymorphisms, and certain polymorphisms may be related to anti-viral treatment response.

Adult ; Alleles ; Antiviral Agents ; therapeutic use ; Cryoglobulinemia ; blood ; complications ; Female ; Genotype ; Hepatitis C, Chronic ; blood ; complications ; drug therapy ; genetics ; Humans ; Interleukins ; genetics ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; RNA, Viral ; blood