OBJECTIVETo establish an improved procedure for isolation and identification of epitopes from a random peptide library displayed on the bacterial surface.

METHODSEpitopes were screened from FliTrx random peptide library by a monoclonal antibody 3B9 against HBV-PreS2 protein. The enrichment was monitored in each round. Higher affinity clones were obtained by increasing the washing strength and randomly selected for sequencing and Western blot analysis.

RESULTSClones specifically binding to antibody were enriched in each round. Ten sequences were obtained from sixteen sequenced clones, seven of them contained the common motif RXRGXY with high homogeneity to 135-140 amio acids in HBV-PreS protein and have positive results in Western blot analysis. The other three sequences have no typical motif RXRGXY and showed different Western blot results.

CONCLUSIONSIt's easy and quick to drive epitopes from a random peptide library displayed on the bacterial surface.

Amino Acid Sequence ; Animals ; Antigens, Viral ; immunology ; Bacterial Proteins ; immunology ; Flagellin ; immunology ; Hepatitis B Surface Antigens ; genetics ; immunology ; isolation & purification ; Immunodominant Epitopes ; immunology ; Molecular Sequence Data ; Peptide Library ; Protein Precursors ; genetics ; immunology ; isolation & purification

OBJECTIVETo study the feasibility of MRI of human colon adenocarcinoma cell line (Lovo) labeled with superparamagnetic iron oxide(SPIO) nanoparticles in vitro.

METHODSLovo cells (5 × 10(5) and 1 × 10(6)) were cultured in medium containing different SPIO nanoparticles (50 microl and 500 microl). Transmission electron microscopy was used to observe cellular ultrastructure and to determine the uptake and distribution of particles in Lovo cells at 1-, 3-, 6-hours. MRI of Lovo cells was performed with T1WI, T2WI sequences. Unlabeled cells were used as controls.

RESULTSUptake of SPIO nanoparticles occurred within 6 hours. On T1 weighted imaging, there was no significant difference in signal intensity between the experimental groups and the control group. On T2 weighted imaging, there was no significant difference in signal intensity between the experimental groups and the control group after culture of 1 h. Signal intensity began to decrease in 1 × 10(6) Lovo cells labeled with 500 microl SPIO nanoparticle after 3 hours culture. Signal intensity decreased in all the experimental groups after 6 hours culture.

CONCLUSIONHuman colon adenocarcinoma cell line (Lovo) can be labeled with SPIO nanoparticles, and the labeled cells can be imaged with MRI equipment.

Adenocarcinoma ; pathology ; Cell Line, Tumor ; Colonic Neoplasms ; pathology ; Humans ; Iron ; Magnetic Resonance Imaging ; methods ; Magnetics ; Nanoparticles ; Oxides ; Pilot Projects ; Staining and Labeling ; methods

[Objective]To establish a reliable and accurate preimplantation genetic diagnosis (PGD)method using multiple dis?placement amplification (MDA), which can be applied to the diagnosis of X-linked severe combined immunodeficiency disease (X-SCID).[Methods]Haplotype analysis for the X-SCID family was performedusing five short tandem repeats (STR) markers flanking the both sides of the interleukin-2 (IL-2) receptor gamma chain (IL2RG) gene. MDA technique was used for single-cell whole genomic amplification. The products were used as template in polymerase chain reaction (PCR) of informative STR markers found by linkage analysis for haplotype analysis as well as sequencing of the IL2RG gene exon 5.The amelogenin (AMEL) locus was used to do sex diag?nosis.[Results]Linked analysis revealed 3 STR markers were informative. The method was evaluated with 10 single lymphocytes and 10 single blastomeres. MDA was successful in all single cell. The detection efficiency of gene sequencing of pathogenic IL 2RG exon5 was 100%. The PCR efficiency of 3 STR informative markers and AMEL was 96.3%(77/80)and the average allele drop-out (ADO) rate was 11.5%(7/61). A cycle of PGD was performed on the family, and seven embryos were diagnosed, two of which were normal embry?os. Twin pregnancy occurred after transplantation which were given a healthy baby boy and a healthy baby girl.[Conclusion]In this study, multiple displacement amplification combined with specific amplification/sequencing of pathogenic gene and haplotype analysis in the single cell level of X-linked severe combined immunodeficiency disease were performed. The protocol can avoid misdiagnosis caused by contamination and ADO, and improve the diagnostic efficiency of PGD.

OBJECTIVETo understand the timeliness of the notifiable communicable diseases surveillance system in Fujian province.

METHODSDatabase from the internet based communicable diseases reporting system was used.

RESULTSThe 50th percentile of time between the disease diagnosed and report recorded in medical faculties was 1 day in 2004 which was 6 days less than that in 2001 - 2003. The timeliness rate of 0 day was 46.46%, a 2.7 times over that in 2001 - 2003. The timeliness of notifiable communicable diseases surveillance system in different administrative areas, reporting units and on different diseases was significantly different. Time between the disease diagnosed and report recorded was the shortest in those cases reported by hospitals and traditional Chinese medicine(TCM) hospitals at the county level and above, with 50th percentile as 0 day, but the timeliness rate of 0 day was 50.76% with 70.04% of the cases were reported from hospitals and TCM hospitals of county level and above. Length between the disease diagnosed and reported was the longest in those cases recorded by Centers for Disease Control and Prevention(CDCs) with the 50th percentile as 3 days. The source of cases recorded by CDCs came from hospitals at the township level, where there was no connection to internet but the reporting cards had to be sent to local CDCs. Time between the disease being diagnosed and reported was 2 days in those cases reported by hospitals at the township level. 21.21% of cases were recorded by hospitals of township level and CDCs. The 50th percentile of time shown between the reported records and confirmed by CDCs was 4 hours The 24 hour timeliness rate was 63.65%.

CONCLUSIONThe timeliness of the notifiable communicable diseases surveillance system had been improved significantly after the medical personnel recording the cases directly through internet. Timeliness could be further improved through access to internet at the hospitals of township level, training of staff and better hospital management systems.

China ; epidemiology ; Communicable Diseases ; epidemiology ; Databases, Factual ; Disease Notification ; methods ; Hospitals ; Internet ; Time Factors

Objective: To investigate the effects of the compound preparation Jinghuosu on oligospermia and asthenospermia. METHODS: This multi-centered clinical study included 120 cases of mild to moderate idiopathic oligospermia or asthenospermia, all treated with oral Jinghuosu once a bag, bid, for 3 successive months. Before and at 1, 2 and 3 months after treatment, we detected sperm concentration, total sperm motility, progressive sperm motility and normal sperm morphology of each ejaculate, and recorded whether the patients had any adverse reactions. RESULTS: After 3 months of treatment, all the patients showed obvious improvement in semen parameters, most significantly in sperm concentration, total sperm motility, and the percentages of progressive motile sperm and morphologically normal sperm (P <0.05). No significant adverse reactions were observed during the 3 months of medication. CONCLUSIONS Jinghuosu has a significant efficacy and no obvious adverse effect in the treatment of mild to moderate oligospermia and asthenospermia.
Asthenozoospermia ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Male ; Oligospermia ; drug therapy ; Semen ; drug effects ; physiology ; Sperm Count ; Sperm Motility

OBJECTIVE: To investigate the incidence rate and risk factors for metabolic bone disease of prematurity (MBDP) in very low birth weight/extremely low birth weight (VLBW/ELBW) infants. METHODS: The medical data of 61 786 neonates from multiple centers of China between September 1, 2013 and August 31, 2016 were retrospectively investigated, including 504 VLBW/ELBW preterm infants who met the inclusion criteria. Among the 504 infants, 108 infants diagnosed with MBDP were enrolled as the MBDP group and the remaining 396 infants were enrolled as the non-MBDP group. The two groups were compared in terms of general information of mothers and preterm infants, major diseases during hospitalization, nutritional support strategies, and other treatment conditions. The multivariate logistic regression analysis was used to investigate the risk factors for MBDP. RESULTS: The incidence rate of MBDP was 19.4% (88/452) in VLBW preterm infants and 38.5% (20/52) in ELBW preterm infants. The incidence rate of MBDP was 21.7% in preterm infants with a gestational age of < 32 weeks and 45.5% in those with a gestational age of < 28 weeks. The univariate analysis showed that compared with the non-MBDP group, the MBDP group had significantly lower gestational age and birth weight, a significantly longer length of hospital stay, and a significantly higher incidence rate of extrauterine growth retardation ( CONCLUSIONS A lower gestational age, hypocalcemia, extrauterine growth retardation at discharge, and neonatal sepsis may be associated an increased risk of MBDP in VLBW/ELBW preterm infants. It is necessary to strengthen perinatal healthcare, avoid premature delivery, improve the awareness of the prevention and treatment of MBDP among neonatal pediatricians, and adopt positive and reasonable nutrition strategies and comprehensive management measures for preterm infants.
Birth Weight ; Bone Diseases, Metabolic/etiology* ; China/epidemiology* ; Female ; Humans ; Infant ; Infant, Extremely Low Birth Weight ; Infant, Newborn ; Infant, Premature ; Infant, Very Low Birth Weight ; Pregnancy ; Retrospective Studies ; Risk Factors

OBJECTIVE: To observe the impact of energy saving light,incandescent light and circadian light on the ethology of depressive rats and explore its possible mechanism on affecting the secretion of melatonin. METHODS: Thirty rats aged 6weeks were randomly selected from 40 specific pathogen free health female SD rats after they adapted to the living environment,depressive rat models were established in the rats by bilateral ovariectomy combined with isolated living and chronic unpredictable mild stress stimulation at the age of 11-14 weeks. Then these 30 ovariectomized rats were randomly divided into 3 intervention groups,including an energy saving light group,an incandescent light group and a circadian light group,with 10 rats in each group. The rats in these 3 groups were given specific experimental light intervention for 3 weeks respectively at the age of 17 weeks. The other 10 rats were raised in conventional environment as the control group. Their body weights were measured at the age of 17,19,20 and 21 weeks. The ethology tests were carried out by sucrose preference test and the open-field test at the age of 7,14 and 20 weeks respectively. The melatonin levels in peripheral blood of 7 time points from 19: 30 to 8: 30 were measured in the rats at age of 21 weeks. One rat in each group at every time point was randomly selected for examination. RESULTS: At the age of 17 weeks before light-intervention,the body weights of rats in 4 groups showed no significant difference( P > 0. 05). After light-intervention,at the age of 17-20 weeks,the body weights of rats in 3 intervention groups were gradually increased with the increase of age( P < 0. 05).There was no significant difference between body weights of rats at the age of 21 weeks and those at the age of 20 weeks in each group( P > 0. 05). At age of 7 weeks,no significant differences were found in sucrose consumption and standing scores among these 4 groups( P > 0. 05). After the depressive models were established,at the age of 14 weeks before light-intervention,in rats of these 3 intervention groups,the sucrose consumption and standing scores were lower than those of the control group( P < 0. 05),and there was no significant difference found in the above 2 indexes among these 3intervention groups( P > 0. 05). At the age of 20 weeks after light-intervention,the sucrose consumption and standing scores were not significantly different from each other among the 4 groups( P > 0. 05). The peak levels of melatonin in the peripheral blood of rats in these 3 intervention groups were higher than that in the control group. The peak levels onsets of melatonin in peripheral blood of rats in the circadian light group and the energy saving light group were earlier or 2 hours delayed compared to that of control group,while it was similar between the incandescent light group and control group.CONCLUSION: The circadian light,the energy saving light and the incandescent light are similarly effective in improving the behaviors of depressive rats. The circadian light can delay the onset of peak level of melatonin in peripheral blood.

Guided by cell-based anti-anaphylactic assay, eighteen cage-like monoterpenoid glycosides (1-18) were obtained from the bioactive fraction of P. lactiflora extract. Among these, compounds 1, 5, 6, 11, 12, 15, and 17 significantly reduced the release rate of β-HEX and HIS without or with less cytotoxicity. Furthermore, the most potent inhibitor benzoylpaeoniflorin (5) was selected as the prioritized compound for the study of action of mechanism, and its anti-anaphylactic activity was medicated by dual-inhibiting HDC and MAPK signal pathway. Moreover, molecular docking simulation explained that benzoylpaeoniflorin (5) blocked the conversion of L-histidine to HIS by occupying the HDC active site. Finally, in vivo on PCA using BALB/c mice, benzoylpaeoniflorin (5) suppressed the IgE-mediated PCA reaction in antigen-challenged mice. These findings indicated that cage-like monoterpenoid glycosides, especially benzoylpaeoniflorin (5), mainly contribute to the anti-anaphylactic activity of P. lactiflora by dual-inhibiting HDC and MAPK signal pathway. Therefore, benzoylpaeoniflorin (5) may be considered as a novel drug candidate for the treatment of anaphylactic diseases.
Animals ; Glucosides ; Mice ; Mice, Inbred BALB C ; Molecular Docking Simulation ; Monoterpenes ; Paeonia ; Plant Roots

Objective: To describe the distribution characteristics of tea consumption in adult twins recruited in the Chinese National Twin Registry (CNTR) and provide clues to genetic and environmental influences on tea consumption. Methods: Enrolled in CNTR during 2010-2018, 25 264 twin pairs aged 18 years and above were included in subsequent analysis. Random effect models were used to estimate tea consumption in the population and regional distribution characteristics. The concordance rate of the behavior and difference in consumption volume of tea within pairs were also described. Results: The mean age of all subjects was (35.38±12.45) years old. The weekly tea consumers accounted for 17.0%, with an average tea consumption of (3.36±2.44) cups per day. The proportion of weekly tea consumers was higher among males, 50-59 years old, southern, urban, educated, and the first-born in the twin pair (P<0.05), and lower among unmarried individuals (P<0.001). Within-pair analysis showed that the concordance rate of tea consumption of monozygotic (MZ) twins was higher than that of dizygotic (DZ) twins and the overall heritability of tea consumption was 13.45% (11.38%-15.51%). Stratified by the characteristics mentioned above, only in males, the concordance rate of MZ showed a tendency to be greater than that of DZ (all P<0.05). The differences in consumption volume of tea within twin pairs were minor in MZ among males (P<0.05), while the differences were not significant in female twins. Conclusion: There were discrepancies in the distribution of tea consumption among twins of different demographic and regional characteristics. Tea consumption was mainly influenced by environmental factors and slightly influenced by genetic factors. The size of genetic factors varied with gender, age, and region, and gender was a potential modified factor.
Adult ; China ; Diet ; Female ; Humans ; Male ; Middle Aged ; Tea ; Twins, Dizygotic ; Twins, Monozygotic ; Young Adult

Objective: To describe the distribution characteristics of type 2 diabetes in twins in Chinese National Twin Registry (CNTR), provide clues and evidence for revealing the influence of genetic and environmental factors for type 2 diabetes. Methods: Of all twins registered in the CNTR during 2010-2018, a total 18 855 twin pairs aged ≥30 years with complete registration information were included in the analysis. The random effect model was used to describe the population and area distribution characteristics and concordance of type 2 diabetes in twin pairs. Results: The mean age of the subjects was (42.8±10.2) years, the study subjects included 10 339 monozygotic (MZ) twin pairs and 8 516 dizygotic (DZ) twin pairs. The self-reported prevalence rate of type 2 diabetes was 2.2% in total population and there was no sighificant difference between MZ and DZ. Intra-twin pairs analysis showed that the concordance rate of type 2 diabetes was 38.2% in MZ twin pairs, and 16.0% in DZ twin pairs, the difference was statistically significant (P<0.001). The concordance rate of type 2 diabetes in MZ twin parts was higher than that in DZ twin pairs in both men and women, in different age groups and in different areas (P<0.05). Further stratified analysis showed that in northern China, only MZ twin pairs less than 60 years old were found to have a higher concordance rate of type 2 diabetes compared with DZ twin pairs (P<0.05). In southern China, the co-prevalence rate in male MZ twin pairs aged ≥60 years was still higher than that in DZ twin pairs (P<0.05). Conclusion: The twin pairs in this study had a lower self-reported prevalence of type 2 diabetes than the general population. The study results suggested that genetic factors play a role in type 2 diabetes prevalence in both men and women, in different age groups and in different areas, however, the effect might vary.
Adult ; China/epidemiology* ; Diabetes Mellitus, Type 2/genetics* ; Diseases in Twins/genetics* ; Female ; Humans ; Male ; Middle Aged ; Registries ; Twins, Dizygotic ; Twins, Monozygotic/genetics*