OBJECTIVEPersistent replication of hepatitis B virus (HBV) is one of the major obstacles in HBV infection treatment. Reduction or clearance of HBV propagation would be one of the aims of HBV therapy. The drugs approved in clinical used such as nucleotide analogs or interferon, were limited effects on HBV replication. The newly developing gene therapy method, dominant negative mutants, were be used as new promising HBV therapy strategy, and a dominant negative mutant of HBVX gene pRev X-GFP which we have reported in our previous study has some effects both on HBV replication and expression in transient expression, but the effects were interfered by persistent secretion of HBV in HepG2 2.2.15 cells without transfection pRev X-GFP in the experiment. To make sure the effects of dominant negative mutant of pRev X-GFP, we established a HBX DN stable express cell clone, and evaluated the effects of HBX dominant negative mutant on HBV replication.

METHODSThe X gene mutant, in which a specific point mutation of 3'-end ATG to AAG and fused with human green fluorescence protein (GFP) were cloned into pRev TRE vector, assigned to pRev HBX-GFP dominant mutant (pRev X-GFP). And the plasmid contains the wild type X gene or GFP gene was cloned into the same vector to construct pRev Xwt, pRev GFP constructs. All the constructs then transfected into HepG2 2.2.15 cells by liposome. After 7 days resistance selection of hygromycin (300 microg/ml), and cell clones which stable expression HBX-GFP, HBXwt, GFP were obtained. After reseeding of 106 cells of each clones in 12 wells with a 12 well cell plate and another 12 wells 2.2.15 cell were serve as blank control. The cells and media were harvested after cultured in DMEM with 10% FBS for 3 days. HBV-related DNA was assayed by dot blot and Southern blot.

RESULTSThe 100% expression of pRev HBX-GFP, GFP and wild type X constructs were obtained. The stable expressed HBX-GFP can significantly reduce HBV DNA level both in cell media and cells by dot blot and Southern blot analysis, but not for pRev Xwt and pRev GFP.

CONCLUSIONThe dominant negative mutant pRev HBX-GFP can significantly inhibit the HBV gene expression. It also suggested that X gene might be one of promising target for HBV gene therapy.

Carcinoma, Hepatocellular ; pathology ; Cloning, Molecular ; DNA Replication ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Hepatitis B virus ; genetics ; physiology ; Humans ; Liver Neoplasms ; pathology ; Point Mutation ; Trans-Activators ; genetics ; Transfection ; Tumor Cells, Cultured ; Virus Replication

Adolescent ; Adult ; China ; Female ; Hepatectomy ; methods ; Humans ; Laparoscopy ; methods ; Liver Transplantation ; methods ; Living Donors ; Male

Allografts ; Hepatectomy ; methods ; Hepatic Veins ; surgery ; Humans ; Iliac Artery ; surgery ; Liver Transplantation ; Middle Aged

Objective: To explore the isotemporal substitution effects among different intensities of physical activity within a 10 minute recess period on the mental health of junior high school students, aiming to provide evidence based references for targeted practical interventions. Methods: From May to November 2024, a total of 845 junior high school students from Tianjin,Taiyuan and L Liang in Shaanxi Province,Puyang in Henan Province,Xi an in Shaanxi Province,Quzhou in Zhejiang Province,and Chaoyang in Liaoning Province were selected by using a combination of stratified random sampling and convenience sampling. ActiGraph wGT3X-BT accelerometers was used to measure physical activity during a 10 minute recess period. Mental health status was assessed with the Depression Anxiety Stress Scale (DASS-21). An isotemporal substitution model was constructed in 1 minute increments to predict the effects of substituting different physical activity behaviors on students mental health. Results: During recess, sedentary behavior (SB) was predominant among junior high school students, with an average duration of [7.08(5.85,7.98)] minutes, while moderate to vigorous physical activity (MVPA) accounted for the shortest duration at [0.42(0.21,0.85)] minutes. There were statistically significant differences in MVPA,LPA and SB time between students of different genders and grades( Z/H =-9.08,-8.34,-9.51;84.87,126.82,135.27,all P <0.01). Isotemporal substitution analysis, adjusted for gender and age, showed that replacing 1 minute of SB with 1 minute of MVPA significantly improved anxiety levels ( β =-0.29, 95% CI =-0.53 to -0.04) and overall mental health ( β =-0.72, 95% CI =-1.39 to -0.04), with both results reaching statistical significance (both P <0.05). No significant effects were observed for other substitution patterns (both P >0.05). Conclusions Substituting SB with MVPA during a 10 minute recess period exerts a positive impact on the mental health of junior high school students. It is recommended to optimize the daily recess activity structure in schools to enhance students mental well being.

OBJECTIVETo investigate and analyze the traumatic conditions of Longnan area of Gansu in Wenchuan earthquake, and to explore the measures coping with unexpected disasters.

METHODSA sample survey including questionnaire survey, interviews and other methods was adopted to investigate the conditions of affected population of Longnan area in earthquake, especially on the traumatic conditions, the injured causes, treatment methods.

RESULTSIn the sample survey of a total of 1652 earthquake casualties, the dead were 16 and the patients with extremities fractures were 750 (45.4%). Organized and timely rescue of the wounded could reduce the complications and the mortality .

CONCLUSIONStrengthening the prevention and treatment of earthquake trauma can effectively decrease the death rate and disabled rate.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; China ; Data Collection ; Disasters ; statistics & numerical data ; Earthquakes ; mortality ; statistics & numerical data ; Female ; Humans ; Male ; Middle Aged ; Wounds and Injuries ; mortality ; psychology ; surgery ; therapy

OBJECTIVETo evaluate the value of 640-slice 3D CT angiography (3D-CTA) plus 3D printing for improving the outcomes of surgeries for intracranial aneurysms.

METHODSSixty patients with intracranial aneurysm were randomly divided into trial group (n=30) and control group (n=30). The control group received routine surgery, and the simulation models of the intracranial aneurysm in trial group was printed using a 3D printer using the imaging data from 3D-CTA. Using the simulation model, the surgery was designed and planned before operation (including surgical approaches and placement of clips) and simulation surgery was also conducted. The coincidence rates between preoperative and intraoperative findings of the intracranial aneurysms on 3D-CTA were compared. CT scan was performed at 1 and 3 days after the operation to detect potential cerebral infarction or bleeding associated with the operation; CTA was performed both at the same time and at 3-6 months after the operation to detect stenosis, occlusion and aneurysm clipping. The patients were followed up for 3-6 months to assess the outcomes using Glasgow Outcome Scale (GOS).

RESULTSThe preoperative 3D-CTA findings were basically consistent with the intraoperative findings in all the 60 patients. Nine patients in the control group and 2 patients in the trial group had short-term adverse operation events; 11 patients in control group and 4 patients in trial group had long-term adverse events; 18 patients in control group and 25 patients in trial group had good neurologic function. The incidences of short-term and long-term adverse events associated with the operation was significantly lower in the trial group than in the control group (χ=5.364, P=0.021; χ=4.841, P=0.028), and the outcomes were significantly better in the trial group than in the control group (χ=4.633, P=0.031).

CONCLUSIONThe simulation model of intracranial aneurysm is helpful to improve the quality of surgery and patients outcomes.

OBJECTIVETo provide evidence for more accurate diagnosis of birth defects based on the pathoanatomy of congenital malformations.

METHODSData used in this study were obtained from Luliang City Hospital and three county hospitals of Shanxi province between February 2004 and March 2006. Autopsy and pathological examination of 160 dead fetuses and stillbirths were performed. Photos of dead fetuses and stillbirths were taken, tissues were cut into sections for pathological examination under microscope, all pathological information was recorded, and percentage of birth defects was calculated.

RESULTSThe proportion of dead fetuses and stillbirths with or without congenital malformations was 84.4% (135/160) and 15.6% (25/160), respectively. There were 16 categories of major external and internal birth defects in 135 cases of such defects. Congenital heart defects, anencephaly and spina bifida had a higher prevalence rate in the study period. The prevalence rate of non-malformation death and birth defects < 28 gestational weeks and internal anomalies > or = 28 gestational weeks was 14.61% (61/4175) and 17.25% (72/4175), respectively. A total of 413 in situ anomalies were found in 135 cases of autopsy. Spina bifida, anencephaly, congenital heart defects, aplasia or accessory lobe of lung, renal agenesis and dysplasis and congenital hydrocephaly were more closely associated with severe malformations than with mitis malformations. The cases of dead fetuses and stillbirths with multiple malformations (> or = 2 in situ anomalies) had a higher proportion (74.1%), whereas those with isolated malformations had a lower proportion (25.93%).

CONCLUSIONThe occurrence of congenital malformations in different embryonic developmental stages affects multiple organs. Postmortem examination of internal and multiple malformations of fetal deaths and stillbirths can provide more accurate diagnostic information for birth defects.

Cause of Death ; China ; epidemiology ; Congenital Abnormalities ; diagnosis ; epidemiology ; Female ; Humans ; Infant, Newborn ; Male ; Population Surveillance ; Pregnancy ; Stillbirth

Animals integrate various environmental stimuli within the nervous system to generate proper behavioral responses. However, the underlying neural circuits and molecular mechanisms are largely unknown. The insulin-like signaling pathway is known to regulate dauer formation, fat metabolism, and longevity in Caenorhabditis elegans (C. Elegans). Here, we show that this highly conserved signaling pathway also functions in the integrative response to an olfactory diacetyl and a gustatory Cu(2+) stimuli. Worms of wild-type N2 Bristol displayed a strong avoidance to the Cu(2+) barrier in the migration pathway to the attractive diacetyl. Mutants of daf-2 (insulin receptor), daf-18 (PTEN lipid phosphatase), pdk-1 (phosphoinositide-dependent kinase), akt-1/-2 (Akt/PKB kinase) and sgk-1 (serum- and glucocorticoid-inducible kinase) show severe defects in the elusion from the Cu(2+). Mutations in DAF-16, a forkhead-type transcriptional factor, suppress the integrative defects of daf-2 and akt-1/-2 mutants. We further report that neither cGMP nor TGFβ pathways, two other dauer formation regulators, likely plays a role in the integrative learning. These results suggest that the insulin-like signaling pathway constitutes an essential component for sensory integration and decision-making behavior plasticity.
Animals ; Caenorhabditis elegans ; genetics ; physiology ; Caenorhabditis elegans Proteins ; genetics ; physiology ; Chemotaxis ; genetics ; physiology ; Copper ; physiology ; Cyclic GMP ; genetics ; physiology ; Diacetyl ; metabolism ; Insulin ; metabolism ; Longevity ; Signal Transduction ; Smell ; genetics ; physiology ; Taste ; genetics ; physiology ; Transforming Growth Factor beta ; genetics ; physiology

OBJECTIVE: To study the application value of multi-slice computed tomography (MSCT) and 3D reconstruction in biomechanical analysis of traffic accidents in forensic medicine. METHODS: Based on a real case, the tomoscan images were obtained from a corpse by MSCT scanning. The 3D images were reconstructed. The biomechanic process of injury manners of impacting, rolling and crushing in traffic accidents was analyzed together with autopsy, vehicle inspection, etc. The MSCT results were compared with the autopsy results. RESULTS: Some characters in situ including the part of fracture on different site that suffered by force from different directions, trends of fracture line, and status of smash inner bones were obtained trough MSCT and 3D reconstruction. Some details like fracture were even better than those from autopsy. CONCLUSION MSCT and 3D reconstruction have some advantages such as in situ reconstruction, easily controlled image and fully conserved evidence. It may be a supplementary method and have a directive function for the biomechanical analysis of traffic accidents.
Accidents, Traffic ; Adult ; Automobiles ; Autopsy/methods* ; Biomechanical Phenomena ; Bone and Bones/injuries* ; Craniocerebral Trauma/pathology* ; Female ; Forensic Pathology ; Fractures, Bone/diagnostic imaging* ; Humans ; Image Processing, Computer-Assisted ; Imaging, Three-Dimensional/methods* ; Tomography, Spiral Computed/methods* ; Wounds and Injuries/pathology*

OBJECTIVETo analyze the cytogenetic characteristics of different age subgroups in patients with acute myeloid leukemia (AML), and to explore the relationship between age and cytogenetics.

METHODSBetween January 2004 and December 2011, Bone marrow (BM) samples from 640 patients with de novo AML were analyzed retrospectively. The analyses were performed according to standard culturing and banding techniques, and clonal abnormalities were defined and described according to the International System for Human Cytogenetic Nomenclature (ISCN 2009). The cytogenetic subtypes were performed as normal, balanced, and unbalanced karyotypes. In the last group, the age distribution of complex and monosome karyotypes were further analyzed. The patients were divided into 8 age groups: 0 - 9, 10 - 19, 20 - 29, 30 - 39, 40 - 49, 50 - 59, 60 - 69, and ≥ 70 year old groups.

RESULTSThe distribution of normal, balanced, and unbalanced karyotypes showed age specific characteristics. The incidence of normal karyotype increased from 6.67% (0 ∼ 9 year old) to 58.33% (≥ 70) (χ(2) = 20.68, P = 0.001) and balanced karyotype decreased from 73.33% (0 ∼ 9) to 11.11% (≥ 70) (χ(2) = 48.22, P < 0.01). The frequency of unbalanced karyotypes increased from 20.0% (0 ∼ 9) to 30.56% (≥ 70) (χ(2) = 18.963, P = 0.008). The frequency of complex karyotype was 6.67% in 0 - 9 year old group, followed by 0% in 10 - 19 and 20 - 29 year old group, and from 1.72% to 11.11% from 30 - 39 to ≥ 70 year old group (χ(2) = 8.341, P = 0.08). Monosome karyotype was only detected in patients in 30 year old or older groups. Although an increased tendency was observed with ages, there was no significant difference (χ(2) = 4.778, P = 0.311).

CONCLUSIONThe different age profiles of the cytogenetic subtypes may indicate the different mechanisms of the pathogenesis of AML, which may also offer beneficial information for etiological research of AML.

Adolescent ; Adult ; Age Factors ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Karyotype ; Karyotyping ; Leukemia, Myeloid, Acute ; genetics ; Male ; Middle Aged ; Retrospective Studies ; Young Adult