HPV16 E7 fusion protein was expressed in E. coli BL21, and its applied value for HPV was evaluated. HPV16 E7 gene was amplified by PCR, and cloned into prokaryotic expression vector pGEX6p-1. The recombinant plasmid was transformed into E. coli BL21, and HPV16 E7 fusion was expressed through IPTG induction. The expressed product was analyzed by SDS-PAGE and Western blot, subsequently purified according to Glutathione Sepharose 4B purification procedure. An indirect ELISA with the purified fusion protein as the coating antigen was then established to detect E7 serum antibodies from mice immunized with recombinant Listeria monocytogenes delivering HPV16 E7. The results demonstrated that the soluble fusion protein was highly expressed at 25 degrees C after induction with 0.5 mM IPTG. Furthermore, the result of Western blot analysis showed that the fusion protein had good specific reaction with an anti-E7 monoclonal antibody. Indirect ELISA result confirmed that the fusion protein could detect the serum antibodies against E7 with a titer of 1:200. The expressed GST-E7 fusion protein was immunocompetent, which was useful in the research of E7 biological function and therapeutic vaccine.
Animals ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; genetics ; Female ; Mice ; Mice, Inbred C57BL ; Papillomavirus E7 Proteins ; biosynthesis ; genetics ; isolation & purification ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis ; isolation & purification

OBJECTIVE: To study the changes of liver phosphofructokinase-2 (PFK-2) level at different postmortem intervals as well as due to different causes of death. METHODS: One hundred and five rats were randomly divided into 3 groups and the rats were sacrificed by either bleeding, suffocating, and neck breaking, respectively. The content of liver PFK-2 at 0, 1, 2, 4, 8, 12 and 24 hours following death were studied using immunohistochemishtry and image analysis. RESULTS: PFK-2 content of the rat's liver in all 3 groups showed a linear decrease at different postmortem intervals with no significant statistical differences found between the different groups. CONCLUSION PFK-2 level in rat liver appears to decrease linearly in correlation with prolonged PMI.
Animals ; Asphyxia/metabolism* ; Cause of Death ; Cervical Vertebrae/injuries* ; Female ; Immunohistochemistry ; Liver/enzymology* ; Male ; Phosphofructokinase-2/metabolism* ; Postmortem Changes ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Shock, Hemorrhagic/metabolism* ; Staining and Labeling ; Time Factors

OBJECTIVETo investigate the characteristic and management of postoperative infection in abdominal cluster transplantation.

METHODSPreliminary experience of two cases of abdominal cluster transplantation including small intestine was reviewed.

RESULTSCombination of five immunosuppressive agents based on tacrolimus was used. Severe Gram-negative bacillus infections occurred. The majority of invasive fungal infections was due to Candida species. Cytomegalovirus (CMV) infection increased monocytes and caused eosinopenia and an inversion of the CD4(+) to CD8(+) cell ratio in recipient I, and human CMV matrix proteins pp71 (CMV-pp71) was detected and identified in bile by PCR. Microabscesses in liver transplant biopsies were presented.

CONCLUSIONSInfectious complications after cluster transplantation were complicated. Strategies to optimize the immunity suppression protocol and early diagnosis and treatment will be important to reduce infection after abdominal cluster transplantation.

Adult ; Bacterial Infections ; drug therapy ; etiology ; Cytomegalovirus Infections ; drug therapy ; etiology ; virology ; Fatal Outcome ; Female ; Humans ; Immunosuppressive Agents ; adverse effects ; therapeutic use ; Intestine, Small ; transplantation ; Liver Transplantation ; adverse effects ; methods ; Male ; Opportunistic Infections ; drug therapy ; etiology ; Organ Transplantation ; adverse effects ; methods ; Postoperative Complications ; drug therapy ; etiology ; Retrospective Studies

Aim To investigate the mechanism by which formononetin (FN) inhibits mitochondrial dynamic-related protein 1 (DRP1) -NLRP3 axis via intervening the generation of ROS to reduce allergic airway inflammation. Methods In order to establish allergic asthma mouse model, 50 BALB/c mice aged 8 weeks were divided into the control group, model group, FN treatment group and dexamethasone group after ovalbumin (OVA) induction. Airway inflammation and collagen deposition were detected by HampE and Masson staining. Th2 cytokines and superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA), and IgE levels in bronchoalveolar lavage fluid (BALF) were measured by ELISA, ROS in BEAS-2B cells was assessed by DCFH-DA staining, DRP1 expression in lung tissue and BEAS-2B cells was detected by immunohistochemistry and immunofluorescence, and the DRP1-NLRP3 pathway was analyzed by immunoblotting. Results FN treatment could effectively ameliorate the symptoms of asthmatic mouse model, including reducing eosinophil accumulation, airway collagen deposition, decreasing Th2 cytokine and IgE levels, reducing ROS and MDA production, increasing SOD and CAT activities, and regulating DRP1-NLRP3 pathway-related protein expression, thereby relieving inflammation. Conclusion FN ameliorates airway inflammation in asthma by regulating DRP1-NLRP3 pathway.

This study aimed to assess the reliability of experienced Chinese orthodontists in evaluating treatment outcome and to determine the correlations between three diagnostic information sources. Sixty-nine experienced Chinese orthodontic specialists each evaluated the outcome of orthodontic treatment of 108 Chinese patients. Three different information sources: study casts (SC), lateral cephalometric X-ray images (LX) and facial photographs (PH) were generated at the end of treatment for 108 patients selected randomly from six orthodontic treatment centers throughout China. Six different assessments of treatment outcome were made by each orthodontist using data from the three information sources separately and in combination. Each assessment included both ranking and grading for each patient. The rankings of each of the 69 judges for the 108 patients were correlated with the rankings of each of the other judges yielding 13 873 Spearman rs values, ranging from -0.08 to +0.85. Of these, 90% were greater than 0.4, showing moderate-to-high consistency among the 69 orthodontists. In the combined evaluations, study casts were the most significant predictive component (R(2)=0.86, P<0.000 1), while the inclusion of lateral cephalometric films and facial photographs also contributed to a more comprehensive assessment (R(2)=0.96, P<0.000 1). Grading scores for SC+LX and SC+PH were highly significantly correlated with those for SC+LX+PH (r(SC+LX)vs.(SC+LX+PH)=0.96, r(SC+PH)vs.(SC+LX+PH)=0.97), showing that either SC+LX or SC+PH is an excellent substitute for all three combined assessment.
Adolescent ; Cephalometry ; standards ; China ; Dental Models ; standards ; Esthetics, Dental ; Female ; Humans ; Male ; Malocclusion, Angle Class I ; therapy ; Malocclusion, Angle Class II ; therapy ; Malocclusion, Angle Class III ; therapy ; Orthodontics ; standards ; Peer Review, Health Care ; standards ; Photography ; standards ; Reproducibility of Results ; Treatment Outcome

Persistent neurogenesis exists in the subventricular zone (SVZ) of the ventricles and the subgranular zone (SGZ) of the dentate gyrus of the hippocampus in the adult mammalian brain. Adult endogenous neurogenesis not only plays an important role in the normal brain function, but also has important significance in the repair and treatment of brain injury or brain diseases. This article reviews the process of adult endogenous neurogenesis and its application in the repair of traumatic brain injury (TBI) or ischemic stroke, and discusses the strategies of activating adult endogenous neurogenesis to repair brain injury and its practical significance in promoting functional recovery after brain injury.
Adult ; Animals ; Humans ; Brain/physiopathology* ; Hippocampus/physiopathology* ; Mammals/physiology* ; Neurogenesis/physiology* ; Brain Hemorrhage, Traumatic/therapy* ; Ischemic Stroke/therapy* ; Recovery of Function ; Spinal Cord/physiopathology*

Objective Identification of the risk factors for extraordinary hidden blood loss (HBL) could clarify the underlying causes and provide more appropriate management. This study aims to identify the predictors of HBL in spinal surgery.Methods Medical records were retrospectively retrieved to collect the data of patients who undergoing posterior thoracic and lumbar fusion surgery or scoliosis surgery. Demographic information, perioperative visible blood loss volume, as well as laboratory results were recorded. The patients receiving fusion surgery or scoliosis surgery were further divided into the HBL positive subgroup and the HBL negative subgroup. Differences in the variables between the groups were then analyzed. Binary logistic regression analysis was performed to determine independent risk factors associated with HBL.Results For patients undergoing posterior spinal surgery, the independent risk factors associated with HBL were autologous transfusion (for fusion surgery P=0.011, OR: 2.627, 95%CI: 1.574-2.782; for scoliosis surgery P<0.001, OR: 2.268, 95%CI: 2.143-2.504) and allogeneic transfusion (for fusion surgeryP<0.001, OR: 6.487, 95%CI: 2.349-17.915; for scoliosis surgery P<0.001, OR: 3.636, 95%CI: 2.389-5.231).Conclusions Intraoperative blood transfusion might be an early-warning indicator for perioperative HBL.
Adolescent ; Adult ; Aged ; Blood Loss, Surgical ; prevention & control ; Blood Transfusion ; Child ; Female ; Humans ; Male ; Middle Aged ; Neurosurgical Procedures ; Retrospective Studies ; Scoliosis ; surgery