This paper was to observe the revaccination effect of domestic yeast-derived recombinant hepatitis B vaccine (YRHBV) to children who were born to HBsAg and HBeAg negative mothers and had been immunized with full course of blood-derived hepatitis B vaccine 4 years ago at their birth. The treated group was revaccinated with 5μg YRHBV while the control group with 10μg blood-derived HB vaccine, their anti-HBs response was determined 1,12,24 months after revaccination. The results showed that a nice anti-HBs response was discovered in group A childeren, no matter how about the anti-HBs level before revaccination was. The kinetics of anti-HBs after YDHBV revaccination was similar to that of the blood-derived HB vaccine, no statistical difference was found. So, 5μg of domestic yeast-derived recombinant HB vaccine gave same immune effect as well as 10μg blood-derived HB vaccine.

Objective To observe the effect of raloxifene, a selective estrogen receptor modulator, on osteoporosis in the osteoprotegerin (OPG) gene knock-out female and male mice. Methods Two groups of OPG gene deficient (OPG-/-) female and male mice, 20 mice in each group, were assigned to raloxifene-treated (3 mg The effect of raloxifene was evaluated by comparing the values of bone mineral density (BMD) , bone strength,histomorphometric measurement and osteoclast number between the raloxifene treated group and placebo group.Results As compared with placebo group osteoporotic manifestations were improved in OPG-/- female mice treated with raloxifene orally. BMD was increased both in lumbar vertebrae (P<0.05) and femurs (P<0.01).Bone strength was measured in femurs by three-point bending test and vertebrae by stress test. Results showed that ultimate load, ultimate stress and Young's modulus were increased both at lumbar and femur bone, suggesting decreased risk of fracture. Tartrate-resistant acid phosphatase, a marker enzyme of osteoclasts, was detected, and the number of osteoclasts declined significantly after the treatment of raloxifene. At the same time, results of histomorphometric measurements indicated that bone trabecular volume was increased and bone formation rate decreased from(8.05±4.02)mm3·mm-2·year-1 to (5.48±1.89)mm3·mm-2· year-1(P＜0.05).These findings were found in the group of OPG-/- female mice treated with reloxifene but not in male mice. Conclusions Raloxifene is effective in treating osteoporosis in female OPG-/- mice, indicating that its action is at least in part independent of OPG gene. But it is ineffective in male OPG-/- mice.

Adiponectin is an adipocyte-derived secretory protein. It was found to be associated with insulin resistance, inflammation and arteriosclerosis. To further study the biological function and expression of adiponection in vivo, adipoenctin gene knock-out and LacZ gene knock-in mouse model was constructed. Gene targeting strategy was designed to replace part of exon 2 and exon 3 of adiponectin gene with full length LacZ gene in frame with remaining upstream ATG and signal peptide sequence of exon 2. The targeting vector (Adipo-LacZ-XpPNT) was constructed and verified by restriction enzyme digestion and sequencing. CJ7 ES cells were transfected with targeting vector linearized by NotⅠ digestion, selected in the medium containing both G418 and ganciclovoir. Resistant clones were screened by PCR and further confirmed by Southern blot for correct homologous recombinants. Chimera mice were obtained by routing microinjection of homologous recombined ES cells into blastocysts. After mating, mice heterozygous and further homozygous for adiponectin knockout and LacZ gene knock-in were established. Expression of both endogenous adiponectin and exogenous LacZ gene in mouse tissues and sera were detected by RT-PCR, Northern-blot, Western blot and ELISA. The results show that adiponectin was disrupted at both mRNA and protein levels. LacZ gene is expressed exclusively in adipose tissue of mutant mice. Its expression profile is identical to endogenous adiponection. Unexpectedly, LacZ activity could not be detected in both adipose tissue and serum although LacZ protein can be detected in adipose tissue but not in serum of mutant mice. In conclusion, mice homozygous for adiponectin knockout and LacZ gene knock-in have been successfully constructed. Mutant mice display LacZ expression profile identical to endogenous adiponectin albeit neither LacZ activity nor protein can be detected in serum of mutant mice.

Objective To analyze the injury types and radiologic findings of the wounded in the major Wenchuan earthquake of May 12, 2008. Methods Radiologic results of 1013 victims admitted to MianYang Central Hospital from May 13, 2008 to May 19, 2008 were studied retrospectively, including X-ray of 854 cases (about 2900 local exam positions) and CT scan of 259 cases (about 300 local exam positions). Results 733 cases showed radiologic abnormalities, including 527 cases of single region injury (accounting for 52. 0%) and 206 cases of multi-region injury (accounting for 20. 3%), the overall positive rate was 72. 4%. The results were in the following order on the basis of the incidence, 288 cases of lower extremity injuries (28.4%), 151 cases of thoracic injuries(14. 9%), 136 cases of upper extremity injuries(13.4%), 133 cases of spine fracture(13.1%), 114 cases of pelvic fracture and dislocation(11.3%),88 cases of craniocerebral injuries (8. 7%), 25 cases of maxillofacial fracture (2. 5%) and 8 cases of abdominal tranma(0. 1%). Thoracic injuries included: rib fracture in 136 cases, pulmonary injury in 59 cases, and sternal fracture in 3 cases. Spinal fracture included: cervical vertebrae involved in 12 cases,thoracic vertebrae involved in 43 cases, lumbar vertebrae involved in 93 cases, and Sacrococcygeal vertebraeinvolved in 12 cases. Craniocerebral injuries included: skull fracture in 41 cases, endocranial abnormalities in 72 cases. Abdominal trauma included: intestinal obstruction in 4 cases, renal contusion in 3 cases and abdominal wall hematoma in 1 case. Conclusion Most parts of the body can be injured by earthquake.Bone fracture and dislocation of lower extremity are the most common manifestations.

OBJECTIVEFactor VIII( FVIII) gene knockout mouse model was established for further study on the treatment of hemophilia A.

METHODSExons 16-19 of the mouse FVIII gene were knocked out by ET clone, ES homologous recombination and tetraploid embryo compensation technology. PCR, reverse transcriptase-PCR(RT-PCR) and immunohistochemistry were used to detect the transcription and translation pattern of FVIII. The phenotype of the knockout mice was analyzed by examining the activated partial thromboplastin time (APTT) and FVIII activity (FVIII:C).

RESULTSPCR, RT-PCR and immunohistochemistry confirmed that FVIII was deficient in the FVIII gene knockout mouse. The APTT results showed that FVIII-deficient mouse plasma had a prolonged clotting time compared to normal mouse plasma. The FVIII:C in heterozygous, hemizygous and homozygous mice was 80%, 8% and 10% of that in normal mice, respectively.

CONCLUSIONThe phenotype of the FVIII gene knockout mouse appears grossly similar to that of human with hemophilia A. Establishment of this model may promote the development of new technologies of treatment to hemophilia A.

Animals ; Disease Models, Animal ; Embryo, Mammalian ; Factor VIII ; genetics ; metabolism ; Female ; Hemophilia A ; genetics ; metabolism ; physiopathology ; Humans ; Male ; Mice ; Mice, Inbred ICR ; Mice, Knockout ; Partial Thromboplastin Time

OBJECTIVETo test whether splicing overlapping extension(SOE) method can be a tool for obtaining rare fusion gene's transcripts and to study the tumorigenic capacity of a novel fusion gene AML1-MTG16.

METHODSSOE method was used to obtain AML1- MTG16 fusion gene's transcripts. MTG16, AML1-MTG16 and AML1-MTG16 without III,VI conserved domains of MTG16 segment were inserted into pEGFP- C1,pDsRed-N1 vector respectively,then transfected NIH3T3 cell line by lipofection. Forty-eight hours later, the transfected cells were examined by laser-scanning confocal microscopy. Stable transfected cells were obtained by G418 500ug/ul selection for one month. Growth curve, soft agar colonies formation tumorigenesis in nude mice were done to compare the difference between stable transfected cells.

RESULTSRecombined AML1-MTG16 by SOE contained its CDS. NIH3T3 expressing AML1-MTG16 had a faster proliferation in medium, colony growth in soft agar. AML1-MTG16 expression cells also induced tumors formation following injection into nude mouse. MTG16,AML1-MTG16 and AML1-MTG16 without III,VI conserved domains of MTG16 were colocalized in the nucleus of cotransfected NIH3T3 cells under the examination of laser-scanning confocal microscope.

CONCLUSIONSOE is an effective method to get rare fusion gene's transcripts. AML1-MTG16 plays an important role in leukemogenesis. MTG16 may also have a carcinogenic property within the AML1-MTG16 fusion gene. Carcinogenic property of AML1-MTG16 is restricted to its localization in the nuclear matrix. N terminal of MTG16 may play an important part in the carcinogenic activity of AML1-MTG16.

3T3 Cells ; transplantation ; Animals ; Cell Division ; genetics ; Cell Transformation, Neoplastic ; genetics ; Cell Transplantation ; Core Binding Factor Alpha 2 Subunit ; Green Fluorescent Proteins ; Luminescent Proteins ; genetics ; metabolism ; Mice ; Mice, Nude ; Microscopy, Confocal ; Neoplasms, Experimental ; genetics ; pathology ; Oncogene Proteins, Fusion ; genetics ; Plasmids ; genetics ; Recombinant Fusion Proteins ; genetics ; metabolism ; Time Factors ; Transcription Factors ; genetics ; Transfection

Objective To investigate the predictive value of neutrophil to lymphocyte ratio (NLR) in peripheral blood for the outcomes in patients with acute intracerebral hemorrhage.Methods Consecutive inpatients with intracerebral hemorrhage diagnosed with the head CT were entolled.The modified Rankin Scale (mRS) was used to evaluate the functional outcomes at 90 d,0-2 wvas defined as good outcome,3-6 were defined as poor outcome,and 6 was defined as death.Univariate analysis was used to compare the demographic characteristics,baseline data,imaging,and laboratory findings between the groups.Multivariate logistic regression analysis was used to determine the independent correlation between NLR and the outcomes,and receiver operating characteristics (ROC) analysis was performed to assess the predictive value of NLR for the outcomes.Results A total of 205 patients with acute intracerebral hemorrhage were enrolled in the study,107 (52.2％) had poor outcome and 57 (27.8％) died.There were significant differences in age (P=0.038),Glasgow Coma Scale (GCS) scores (P=0.001),National Institutes of Health Stroke Scale (NIHSS) scores (P =0.001),neutrophil count (P =0.005),lymphocyte count (P =0.002),NLR (P =0.001),fasting blood glucose (P =0.012),hypersensitivity C-reactive protein (P=0.002),hematoma volume (P =0.005),and proportion of bleeding into the ventricles (P =0.002) between the poor outcome group and the good outcome group.There were significant differences in age (P =0.002),previous stroke (P =0.018),GCS scores (P =0.001),NIHSS scores (P =0.001),neutrophil count (P=0.008),lymphocyte count (P=0.001),NLR (P=0.001),fasting blood glucose (P=0.016),hematoma volume (P=0.001),and proportion of bleeding into ventricle (P=0.002) between the death group and the survival group.Multivariate logistic regression analysis showed that NLR was an independent predictive factor for poor outcome (odds ratio [OR] 2.405,95％ confidence interval [CI] 1.613-3.587;P=0.001) and death (OR 2.268,95％ CI 1.532-3.358;P =0.001) after adjusting for confounders.The ROC curve analysis showed that NLR had a higher predictive value for poor outcome at 90 d (area under the ROC curve 0.703,95％ CI 0.632-0.774;P ＜ 0.001).When the cutoff value was 2.3,the sensitivity and specificity were 61.7％ and 72.4％,respectively.NLR also had a predictive value for death within 90 d (area under the ROC curve 0.706,95％ CI 0.629-0.786;P =0.003).When the cutoff value was 2.2,the sensitivity and specificity were 63.2％ and 72.6％,respectively.Conclusion NLR may have certain predict value for outcomes in patients with acute intracerebral hemorrhage.

Objective:To analyze the diagnostic and prognostic value of routine bone marrow examination in patients with extranodal NK/T-cell lymphoma (ENKTCL) based on PET-CT staging.Methods:Clinical data of 186 patients who received bone marrow biopsy and bone marrow aspiration in Fujian Medical University Union Hospital from 2013 to 2021 were retrospectively analyzed. All patients were divided into bone marrow biopsy + bone marrow aspiration group ( n=186) and PET-CT + bone marrow biopsy group ( n=139). The sensitivity, specificity, positive and negative predictive values were compared between two groups. The data were analyzed and plotted. Survival analysis was performed using Kaplan-Meier method and log-rank test. Results:In the whole cohort, 45 patients were positive for bone marrow biopsy, and 30 of them were positive for bone marrow aspiration. A total of 141 patients who were negative for bone marrow biopsy also achieved negative results for bone marrow aspiration. A total of 139 patients completed PET-CT staging and bone marrow biopsy. And 30 patients were diagnosed with positive bone marrow by PET-CT, in which 22 of them were confirmed positive by bone marrow biopsy. Among 109 patients diagnosed with negative bone marrow by PET-CT, 5 of them were confirmed positive by bone marrow biopsy. All these cases were classified as stage Ⅳ due to distant metastases. PET-CT had a diagnostic sensitivity of 81.5%, a specificity of 92.9%, a positive predictive value of 73.3%, and a negative predictive value of 95.4%. Among early stage (Ⅰ-Ⅱ stage) patients diagnosed with PET-CT, all of them were negative for bone marrow biopsy (the negative predictive value was 100%). In stage Ⅳ patients ( n=55), the 1-year overall survival of patients with bone marrow involvement by bone marrow biopsy or PET-CT ( n=35) compared with their counterparts with the involvement of other organs ( n=20) was 28.7% vs.42.0% ( P=0.13), and 1-year progression free survival rates was 23.2% vs. 23.3% in ( P=0.94). Conclusions:Routine bone marrow biopsy does not change the original staging of patients with early stage ENKTCL based on PET-CT staging. Advanced stage patients with positive bone marrow biopsy tend to obtain worse prognosis, indicating that bone marrow biopsy still has certain value.