BACKGROUND: Recently, one focus of research has been Annexin Ⅴ (AnV) existing on hepatic cells membranes as a fundamental receptor related to hepatitis B virus (HBV) infection. Also its expression in placental tissues has been a matter of debate. The study of the relationships between placental cells infected with HBV and their AnV expression will be of great value in future prevention strategies and treatments.OBJECTIVE: To investigate the presence of AnV in HBV infected human's placental cells and its potential role in HBV intrauterine transmission.DESIGN: Randomized controlled experiment.SETTING: Taishan Medical College.MATERIALS: Placental tissue was collected from HBsAg positive full term pregnant women (30 cases) admitted to Jinan Institute for Maternal and Child Health, Taian Central Hospital and Taian Institute for Maternal and Child Health from January 2003 to December 2004. Maternal serum was also obtained. Informed consents for participating in this study were obtained from all the involved pregnant women and this experiment was approved by the Hospital Ethics Committee. Rabbit-anti-human AnV purified affinity antibody (first antibody), rat-anti-human HBs mAb (first antibody),and biotinylated goat-anti-mouse IgG (secondary antibody) were supplied by Wuhan Boster Bioengineering Company.METHODS: Using SABC immunohistochemical staining reagent, 18 HBsAg positive placentas were obtained from 30HBsAg infected patients in full term pregnancy. These were considered as the positive group and the other 12 were used as negative controls. The staining process included dewaxing, dehydration of embedded slides and microwave antigen restoration. In the wet box, rabbit-anti-human AnV purified antibody (first antibody, 1:60, monoclonal antibody)was added on the slides and kept at 4 ℃ overnight. Rat-anti-human antibody HBs mAb(secondary antibody, 1:50) was added and kept at 4 ℃ ovemight, after this procedure, biotinylated goat-anti-mouse IgG(1:100), the first fluorescent antibody such as FITC-goat anti-rabbit IgG (1:50) and the second fluorescent antibody (Avidin-Cy3) were used,respectively. The slides were sealed with buffered glycerol and examined under a confocal laser scanning microscope.The images on the slides were analyzed with IPP 4.5 image programs.MAIN OUTCOME MEASURES: Detecting the simultaneous existence and distribution of HBsAg/AnV in placental cells with HBV infection.RESULTS: Ten cases from the positive group were simultaneously detected for HBsAg/AnV by double-labeled immunofluorenscence assay and confocal laser scanning microscope. AnV expression was detected in the trophoblastic, interstitial cells and vascular endothelial cells of villi interstitial blood vessels, and the coexistence of HBsAg/AnV was found even in one cell.CONCLUSION: HBsAg combined with the receptor AnV in the same placental cells is a common finding in HBV infected full term pregnant women. This finding is very suggestive of a mechanism where AnV could promote hepatitis B virus to enter the placental cells and cause intrauterine infection.

The recombinant human Smad7 adenoviral vector was constructed by direct DNA cloning protocol and then transfected into 293 cells for virus packaging. After amplification and purification, the recombinant adenovirus was used to infect the keloid fibroblasts. The Smad7 mRNA transcription of the infected cells was detected by RT-PCR. The recombinant Adeno-Smad7 was correctly constructed and confirmed by both restriction analysis and PCR analysis. RT-PCR showed the over expression of adenovirus mediated Smad7 mRNA in keloid cells. These results demonstrated that the recombinant Smad7 adenoviral vector can be expressed in cultured cells in vitro, and it may provide a new therapeutic strategy for keloid gene therapy.
Adenoviridae ; genetics ; metabolism ; Cells, Cultured ; Fibroblasts ; metabolism ; pathology ; Genetic Vectors ; genetics ; metabolism ; Humans ; Keloid ; metabolism ; pathology ; RNA, Messenger ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Smad7 Protein ; biosynthesis ; genetics ; Transfection

OBJECTIVETo study transforming growth factor-beta1 (TGF-beta1) autoproduction in keloid fibroblasts and the regulation effect of blocking TGF-beta intracellular signaling on rhTGF-beta1 autoproduction.

METHODSKeloid fibroblasts cultured in vitro were treated with either rhTGF-beta1 (5 ng/ml) or recombinant adenovirus containing a truncated type II TGF-beta receptor gene (50 pfu/cell). Their effects of regulating gene expression of TGF-beta1 and its receptor I and II were observed with Northern blot.

RESULTSrhTGF-beta1 up-regulated the gene expression of TGF-beta1 and receptor I, but not receptor II. Over-expression of the truncated receptor II down-regulated the gene expression of TGF-beta1 and its receptor I, but not receptor II.

CONCLUSIONSTGF-beta1 autoproduction was observed in keloid fibroblasts. Over-expression of the truncated TGFbeta receptor II decreased TGF-beta1 autoproduction via blocking TGF-beta receptor signaling.

Activin Receptors, Type I ; biosynthesis ; pharmacology ; Cells, Cultured ; Down-Regulation ; Fibroblasts ; drug effects ; metabolism ; Gene Expression ; Humans ; Keloid ; metabolism ; Protein-Serine-Threonine Kinases ; RNA, Messenger ; genetics ; metabolism ; Receptors, Transforming Growth Factor beta ; biosynthesis ; metabolism ; Sensitivity and Specificity ; Signal Transduction ; Trans-Activators ; metabolism ; Up-Regulation

Objective: To monitor the antimicrobial resistance and drug-resistance genes of Yersinia enterocolitis, Y. intermedia and Y. frederiksenii recovered from retailed fresh poultry of 4 provinces of China. Methods: The susceptibility of 25 isolated Yersinia spp. to 14 classes and 25 kinds of antibiotics was determined by broth microdilution method according to CLSI (Clinical and Laboratory Standards Institute). The antibiotic resistance genes were predicted with antibiotic resistance genes database (ARDB) using whole genome sequences of Yersinia spp. Results: In all 22 Y. enterocolitis tested, 63.7% (14 isolates), 22.8% (5 isolates), 4.6% and 4.6% of 1 isolates exhibited the resistance to cefoxitin, ampicillin-sulbactam, nitrofurantoin and trimethoprim-sulfamethoxazole, respectively. All the 25 isolates were multi-drug resistant to more than 3 antibiotics, while 64.0% of isolates were resistant to more than 4 antibiotics. A few Y. enterocolitis isolates of this study were intermediate to ceftriaxone and ciprofloxacin. Most Yersinia spp. isolates contained antibiotic resistance genes mdtG, ksgA, bacA, blaA, rosAB and acrB, and 5 isolates recovered from fresh chicken also contained dfrA1, catB2 and ant3ia. Conclusion The multi-drug resistant Yersinia spp. isolated from retailed fresh poultry is very serious in the 4 provinces of China, and their contained many kinds of drug-resistance genes.

Objective:To describe the trend of mortality and death spectrum in Shandong Province from 1970 to 2021 and provide basis for the targeted disease prevention and control.Methods:The data were collected from the death registration reports of Shandong and 3 national retrospective surveys of death causes in Shandong. The change in levels of overall and specific deaths in Shandong in different years were analyzed based on mortality rate, age-standardized mortality rate and constituent ratio of cause of death, differential decomposition was used to quantify the contribution of demographic and non-demographic factors to changes of mortality.Results:The crude mortality rate in residents in Shandong was basically stable from 1970 to 2021, and the mortality rate during 2020-2021 (732.73/100 000) was slightly higher than that during 1970-1974 (671.98/100 000). While the standardized mortality rate decreased significantly, and the mortality during 2020-2021 (183.39/100 000) decreased by 67.71% compared with that during 1970-1974 (568.00/100 000). The negative increase of population factors and the positive decrease of non-population factors reacted each other, so the mortality was relatively stable. Cardiac-cerebrovascular disease was always the leading cause of death, but the constituent ratio of death increased rapidly from 19.70% during 1970-1974 to 54.72% during 2020-2021. The rank in the causes of death changed from the fourth (11.46%) to the second (25.70%) for malignant tumor, from the seventh (5.85%) to the third (5.59%) for injury, from the second (12.87%) to the fourth (4.99%) for chronic respiratory diseases, from the third (12.27%) to the tenth (0.42%) for infectious diseases. The standardized mortality rates of the main causes of death decreased at different degrees, the standardized mortality rates of obstetrical disease, infectious disease, gastrointestinal disease and chronic respiratory disease decreased by more than 50.00%. The age distribution of deaths and the death spectrum in different age groups and in urban-rural populations changed significantly. During 2020-2021, the proportion of deaths in young people aged 0-14 years was 0.54%, which was 97.05% lower than that during 1970-1974, while the proportion of deaths in the elderly aged ≥75 years was 55.14%, which was 55.75% higher than that during 1970-1974. The rank of infectious diseases in the causes of death descended significantly in all age groups, but the ranks of injury, neuropsychiatric disease and malignant tumor rose significantly in adolescents, and the ranks of endocrine nutrition and metabolic disease rose in middle-aged and elderly people. The difference of death spectrum between urban area and rural area became less obvious and the main death causes in urban and rural residents were basically the same during 2020-2021.Conclusions:The death spectrum of residents in Shandong changed significantly. Chronic and non-communicable diseases, especially cardiac-cerebrovascular disease and malignant tumor, should be the focus in disease control and prevention. The prevention and control of diseases in Shandong made remarkable achievement during 1970-2021. However, in the context of population ageing, it is suggested to strengthen the treatment, prevention of diseases and injuries related to the health of the elderly and elderly health care in the future.

Objective To investigate the clinical characteristics of 8 immunodeficiency cases caused by human recombination activating gene 1 (RAG1) mutations,and to explore the relationship among genotypes,clinical manifestations and immunophenotypes.Methods Clinical data were collected and analyzed from patients with RAG1 mutations who visited the Department of Clinical Immunology,Children's Hospital of Fudan University between October 2013 and June 2017.The data included clinical manifestations,immunophenotypes and genotypes.Results A total of 8 patients were diagnosed with RAG1 deficiency (6 boys and 2 girls).The minimum age of onset was 2 months,and the maximum age was 4 months.The minimum age of diagnosis was 2 months,and the maximum age was 13 years.Four patients had a family history of infant death due to severe infections.Two cases were born to the same consanguineous parents.All cases had recurrent infections,including involvement of respiratory tract (8 cases),digestive tract (6 cases),urinary tract (1 case),and central nervous system (1 case).The pathogens of infection included bacteria,viruses and fungi.Rotavirus was found in 3 cases,cytomegalovirus (CMV) in 5 cases,bacillus Calmette-Guérin adverse reaction in 2 cases (1 of whom had a positive acid-fast smear from lymph node puncture fluid),fungal infection in 3 cases.One case had multiple nodular space-occupying lesions in lungs and abdominal cavity complicated with multiple bone destruction.The peripheral blood lymphocyte counts of all patients ranged between 0.1 × 109/L and 3.3 × 109/L (median,0.65 × 109/L).Eosinophilia was found in 3 cases (range,(0.48-1.69) x 109/L).The patients were classified according to immunophenotype as severe combined immunodeficiency phenotype (4 cases),leaky severe combined immunodeficiency (2 cases),Omenn syndrome (1 case) and combined immunodeficiency(1 case).Decreased serum IgG levels were found in 3 cases,increased serum IgM levels in 3 cases,increased serum IgE levels in 5 cases.RAG1 homozygous mutations were detected in 5 cases and RAG1 compound heterozygous mutations in 3 cases.Two novel mutations and six previously reported mutations were identified.Three cases were successfully treated with hematopoietic stem cell transplantation.Four cases died due to infections,and the 13 year-old patient was still under follow-up in the outpatient clinic.Conclusions Different RAG1 gene mutations can lead to diverse clinical presentations and immune phenotypes.Clinicians should pay attention to the family history of infant death with severe infection.In that situation,immunological evaluation and gene detection should be performed as early as possible.