In order to develop monoclonal antibodies (McAbs) against the gp90 protein of reticuloendotheliosis virus (REV), the His-tagged gp90 protein of REV was used to immunize BALB/c mice. Hybridomas were generated by fusing mouse myeloma cells SP2/0 with the splenocytes from the immunized mice. After screening and 3 rounds of cloning process, 3 hybridomas (3G5-B8, 3G5-A10 and 1G12) that stably secreted McAbs against the REV-gp90 were obtained. The isotypes of the McAbs were determined to be IgG1, IgG1 and IgG2b. The McAbs specifically bound to gp90 in REV-infected DF-1 cells, as demonstrated by Western blotting and indirect immunofluorescence assay. The recognition regions on gp90 that were recognized by 3G5-B8/3G5-A10 and 1G12 were located between amino acids 200 to 245 and 230 to 235, respectively, as demonstrated by Western blotting analysis. These McAbs will be useful in the diagnosis and pathogenesis study of REV.
Animals ; Antibodies, Monoclonal ; biosynthesis ; Antibody Specificity ; Blotting, Western ; Epitope Mapping ; Hybridomas ; Immunoglobulin G ; biosynthesis ; Mice ; Mice, Inbred BALB C ; Reticuloendotheliosis virus ; immunology ; Viral Envelope Proteins ; immunology

Infectious bursal disease virus (IBDV) VP4 plays an important role in immunosuppression of host. In order to develop monoclonal antibodies (McAbs) against VP4, we vaccinated BALB/c mice with His-VP4, screened and subcloned positive clones. We established 4 hybridoma cell lines that stably secreted McAbs against VP4 and named these cell lines 3B3, 3H11, 4C8 and 4G6, respectively. We tested the dissociation constant (Kd) of these McAbs, and found that their K(d)s were 4.61 x 10(-11), 1.71 x 10(-10), 4.26 x 10(-11), 5.02 x 10(-11), respectively. The isotypes of these McAbs were determined to be IgG1, IgG1, IgG2b and IgG1. These McAbs specifically bound to VP4 in IBDV infected DF-1 cells as demonstrated by Western blotting analysis and fluorescence antibody assay. These McAbs would help to detect IBDV infection and to analyze the biological activities of IBDV VP4.
Animals ; Antibodies, Monoclonal ; biosynthesis ; Blotting, Western ; Cell Line ; Fluorescent Antibody Technique ; Hybridomas ; Infectious bursal disease virus ; Mice ; Mice, Inbred BALB C ; Viral Structural Proteins ; immunology

Objective: To investigate the effects of hepatitis B virus (HBV) genotype and mutations on the development of hepatocellular carcinoma (HCC) and to establish a new qualified HCC risk scores. Methods: A cohort study enrolling patients with chronic HBV infection was conducted. HBV genotypes were identified by nested multiplex PCR. HBV mutations in the basic core promoter region and PreS region were sequenced after PCR amplification. Scores on risk factors were set based on nomogram. Results: Totally, 1 525 patients were followed-up in this research. A total of 1 110 patients infected with genotype C were followed-up for 8.52 (Q(R): 5.36-11.68) years on average, of whom the incidence of HCC was 11.93/1 000 person-years. In genotype C HBV infected patients, male gender, aged 40 years and over, and four DNA mutations (T1674CG, A1762T/G1764A, A3120T, and A2962G) can increase the risk of HCC (P<0.05); interferon therapy can reduce the risk of HCC (P<0.05). A new HCC predicting model was established according to the results. After validation, the predicted disease-free survival rate was consistent with the real one. Conclusions: Hepatitis B virus genotypes and mutations were closely associated with HCC. The new risk scoring system can well predict HCC occurrence in genotype C HBV infected patients.
Adult ; Aged ; Carcinoma, Hepatocellular/virology* ; China/epidemiology* ; Cohort Studies ; DNA, Viral/genetics* ; Female ; Genotype ; Hepatitis B virus/genetics* ; Hepatitis B, Chronic/complications* ; Humans ; Liver Neoplasms/virology* ; Male ; Middle Aged ; Mutation ; Predictive Value of Tests ; Risk Factors ; Sensitivity and Specificity

When facing the worldwide abuse of opioid substance, one of the effective responses is opioid substitution therapy (OST). However, different OST service patterns may affect the therapeutic outcome. Using the System Engineering Initiative for Patient Safety (SEIPS) model, we can analyze the factors that affecting the outcomes of patients from the perspective work system. In this paper, SEIPS model is used to describe the existing OST service model. According to the operation mechanism of the methadone maintenance treatment in China and the existing OST service model, some suggestions are put forward to carry out effective OST service in the country.
Analgesics, Opioid/adverse effects* ; China ; Delivery of Health Care ; Humans ; Methadone/therapeutic use* ; Opiate Substitution Treatment ; Opioid-Related Disorders/therapy* ; Treatment Outcome

The bones are the most common sites of breast cancer metastasis. Upon arrival within the bone microenvironment, breast cancer cells coordinate the activities of stromal cells, resulting in an increase in osteoclast activity and bone matrix degradation. In late stages of bone metastasis, breast cancer cells induce apoptosis in osteoblasts, which further exacerbates bone loss. However, in early stages, breast cancer cells induce osteoblasts to secrete inflammatory cytokines purported to drive tumor progression. To more thoroughly evaluate the role of osteoblasts in early stages of breast cancer metastasis to the bones, we used green fluorescent protein-labeled human breast cancer cell lines MDA-MB-231 and MDA-MB-435, which both induce osteolysis after intra-femoral injection in athymic mice, and the murine pre-osteoblastic cell line MC3T3-E1 to modulate osteoblast populations at the sites of breast cancer metastasis. Breast cancer cells were injected directly into the femur with or without equal numbers of MC3T3-E1 cells. Tumors grew significantly larger when co-injected with breast cancer cells and MC3T3-E1 cells than injected with breast cancer cells alone. Osteolysis was induced in both groups, indicating that MC3T3-E1 cells did not block the ability of breast cancer cells to cause bone destruction. MC3T3-E1 cells promoted tumor growth out of the bone into the extraosseous stroma. These data suggest that breast cancer cells and osteoblasts communicate during early stages of bone metastasis and promote tumor growth.
Animals ; Bone Neoplasms ; secondary ; Breast Neoplasms ; pathology ; Cell Line ; Cell Line, Tumor ; Disease Models, Animal ; Female ; Femur ; pathology ; Humans ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Osteoblasts ; cytology ; physiology ; Osteolysis ; etiology ; Tumor Burden

Objective: To explore the sampling method in China National Human Biomonitoring Program (HBP) and the related errors, so as to calculate and evaluate the study design in sampling. Methods: The sampling method of HBP is of multistage nature. Taking the results of sampling method from Guizhou province as an example, results related to sampling error and variation coefficient were calculated, using the multistage unequal probability sampling error method. Results: The HBP covered 152 monitoring sites in 31 provinces (autonomous regions and municipalities) and with 21 888 residents selected. The replacement rates at various stages were 5.26%, 6.35% and 40.6% respectively. The sampling error in Guizhou province was 3 207 594, and the coefficient of variation was 0.097. Conclusions: According to the multi-stage unequal probability sampling method, the sampling coefficient variability appeared small with high precision, in Guizhou province. However, this method did not consider the weight adjustment of non-sampling errors such as population missing rate and response rate. Methods related to the calculation on multi-stage sampling error among large-scale public health monitoring projects need to be further studied.
China ; Cities ; Environmental Monitoring ; Humans ; Research Design

OBJECTIVETo observe two-year natural progression of chronic periodontitis in mild, moderate and advanced periodontitis patients.

METHODSThe periodontal status of 169 untreated chronic periodontitis patients aged from 22 to 64, were examined for two years. Periodontal measurements were performed on all teeth except the third molars and 6 sites examined for each tooth. Probing depth (PD), attachment loss (AL), and bleeding on probing (BOP) were measured at baseline, one year, and two year by a same experienced periodontist. Forty-five patients were diagnosed as having mild periodontitis, 87 with moderate, and 37 with advanced periodontitis. The changes of attachment level in these three group patients were analyzed. The site with change of AL greater than 3 mm (DeltaAL > or = 3 mm) were defined as periodontal disease activity (PDA) sites. The occurrence of PDA in three groups was compared.

RESULTS(1) The average AL levels at 1 year and at 2 year were greater than that at baseline in mild, moderate and advanced periodontitis. (2) The percentage of sites with AL > or = 1 mm in three groups all increased from baseline to 1 year and to 2 year. (3) The occurrence of periodontal disease activity increased significantly from mild (0.14% at site level, 15.56% at subject level), moderate (0.39%, 29.89%) to advanced (0.73%, 43.24%) periodontitis patients. (4) The mean baseline AL and PD levels in active sites were greater than that in inactive sites (PD: 3.03 +/- 0.45 vs. 2.87 +/- 0.38, P < 0.05; AL: 2.25 +/- 0.93 vs. 1.77 +/- 0.90, P < 0.01).

CONCLUSIONUntreated advanced periodontitis patients were the risk population for further periodontal breakdown.

Adult ; Chronic Disease ; Disease Progression ; Female ; Humans ; Longitudinal Studies ; Male ; Middle Aged ; Periodontal Attachment Loss ; diagnosis ; Periodontal Index ; Periodontitis ; diagnosis ; Prospective Studies

Epidemiology is a traditional subject mainly based on principles and concepts, and its teaching method needs further improving to meet the requirement of the new trend of education reform. Lecture-based teaching, problem-based teaching, case-based teaching, and internet based teaching, such as flip class, massive open online course and micro-lecture, all have its own unique merits in the practice of epidemiology teaching. So the combination of traditional teaching and online teaching is the most promising mode. "Rain class" , a mixed mode, is an efficient tool to present the epidemiology case more actually in class. Thus, teaching design and application of "rain class" are worth research.
Computer-Assisted Instruction ; Epidemiology/education* ; Humans ; Internet ; Online Systems ; Research ; Teaching

A 15-day-old boy was admitted to the hospital due to repeated convulsions for 14 days. The main clinical manifestations were uncontrolled seizures, hypoergia, feeding difficulties, limb hypotonia, and bilateral hearing impairment. Clinical neurophysiology showed reduced brainstem auditory evoked potential on both sides and burst-suppression pattern on electroencephalogram. Measurement of very-long-chain fatty acids in serum showed that C26:0 was significantly increased. Genetic testing showed a pathogenic compound heterozygous mutation, c.101C>T(p.Ala34Val) and c.1448_1460del(p.Ala483Aspfs*37), in the
Genetic Testing ; Humans ; Infant, Newborn ; Male ; Muscle Hypotonia ; Mutation ; Peroxisomal Multifunctional Protein-2/genetics* ; Protein Deficiency/genetics*

This report presents a case of a male infant, aged 32 days, who was admitted to the hospital due to 2 days of bloody stools and 1 day of fever. Upon admission, venous blood samples were collected, which appeared pink. Blood biochemistry tests revealed elevated levels of triglycerides and total cholesterol. The familial whole genome sequencing revealed a compound heterozygous variation in the LPL gene, with one variation inherited from the father and the other from the mother. The patient was diagnosed with lipoprotein lipase deficiency-related hyperlipoproteinemia. Acute symptoms including bloody stools, fever, and bloody ascites led to the consideration of acute pancreatitis, and the treatment involved fasting, plasma exchange, and whole blood exchange. Following the definitive diagnosis based on the genetic results, the patient was given a low-fat diet and received treatment with fat-soluble vitamins and trace elements, as well as adjustments to the feeding plan. After a 4-week hospitalization, the patient's condition improved and he was discharged. Follow-up showed a decrease in triglycerides and total cholesterol levels. At the age of 1 year, the patient's growth and psychomotor development were normal. This article emphasizes the multidisciplinary diagnosis and treatment of familial hyperlipoproteinemia presenting with symptoms suggestive of acute pancreatitis, including bloody ascites, in the neonatal period.
Humans ; Infant ; Male ; Acute Disease ; Ascites ; Cholesterol ; Hyperlipoproteinemia Type I/genetics* ; Hyperlipoproteinemias ; Lipoprotein Lipase/genetics* ; Pancreatitis ; Triglycerides