Previously the diseases of pediatric hyperandrogenism were mainly diagnosed and evaluated by testing traditional androgens such as testosterone and androstenedione.However, clinical application has revealed a poor correlation between traditional androgens and the clinical manifestations of hyperandrogenism in some patients.It has been proposed that adrenal-derived 11-oxygenated androgen may also be involved in the course of this type of disease.The concentrations of 11-oxygenated androgen are elevated in androgen excess diseases, and they fulfill a variety of roles in human physiology and disease.This article discusses three aspects of the synthesis process, activity and content of 11-oxygenated androgen and their application in three androgen excess diseases: congenital adrenocortical hyperplasia, premature adrenarche and polycystic ovary syndrome, in order to help clinicians expand their clinical understanding and investigative thoughts on 11-oxygenated androgen.

OBJECTIVETo investigate the high expression of HPV16L2N120E7E6 fusion protein by prokaryotic expression system, and evaluate its immunogenicity and antitumor efficacy in vaccinated mice.

METHODSThe HPV16L2N120E7E6 fusion gene, its codons were optimized to increase the expression of the protein, was constructed by overlap extension PCR and inserted into prokaryotic expression vector pET9a. Then the fusion protein was expressed by inducing with IPTG in E. coli strain BL21 (DE3) harboring with plasmid pETL2N120E7E6, and further detected by SDS-PAGE and Western-blot. Finally, the humoral and cellular immune responses were measured by ELISA and ELISPOT, respectively, in vaccinated mice with the purified HPV16L2N120E7E6 fusion protein, and the antitumor efficacy was assessed in mice using the TC-1 tumor challenge model.

RESULTSThe codon-optimized HPV16L2N120E7E6 fusion gene was highly expressed in E. coli strain BL21 (DE3) harboring with plasmid pETL2N120E7E6, and the amount of fusion protein was nearly 48.6% of the total bacterial protein. The purified fusion protein could induce high titer of specific antibody against L2, E7 and E6 in vaccinated mice. When accompanied with the adjuvant CpG, the fusion protein was able to elicit strong and moderate cellular immune responses in vaccinated mice against peptide HPV16E7(49-57) and peptide pools of HPV16E6, respectively. Furthermore, the tumor therapeutic experiment showed that HPV16L2N120E7E6 + CpG could prevent the tumor formation in 80.0% (8/10) vaccinated mice.

CONCLUSIONSThe data of this study suggest that HPV16L2N120E7E6 fusion protein could be a promising candidate vaccine for treatment of chronic HPV16 infection and post-operative adjuvant therapy for cervical cancer.

Adjuvants, Immunologic ; pharmacology ; Animals ; Cancer Vaccines ; immunology ; therapeutic use ; Capsid Proteins ; genetics ; immunology ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Codon ; Escherichia coli ; immunology ; metabolism ; Female ; Humans ; Immunization ; methods ; Immunotherapy ; methods ; Mice ; Mice, Inbred C57BL ; Neoplasm Transplantation ; Oligodeoxyribonucleotides ; immunology ; Oncogene Proteins, Viral ; genetics ; immunology ; metabolism ; Papillomavirus E7 Proteins ; genetics ; immunology ; metabolism ; Papillomavirus Vaccines ; immunology ; therapeutic use ; Plasmids ; Recombinant Fusion Proteins ; genetics ; immunology ; metabolism ; Repressor Proteins ; genetics ; immunology ; metabolism

Objective To determine the changes of cerebrospinal fluid(CSF) ?-endorphin(?-EP) and C-reactive protein(CRP) in children with central nervous system(CNS) infection.Methods Sixty-five children suffered from CNS infection were determined the plasma and CSF ?-EP and CRP concentration during the acute and recovering stage with radioimmunoassay, which included 48 viral encephalitis, 12 purulent meningitis and 5 tuberculou meningitis,and 24 non-CNS disease children were as control group.Results The concentrations of plasma and CSF ?-EP of every experimental group were obviously higher than those of control group during the early stage of CNS infection and these were obviously lower during the recovering stage. The serum concentration of CRP during acute stage was significantly higher than that during recovering stage. No change of serum and CSF CRP concentration was determined during either the acute or recovering stage in the other two experimental groups.Conclusions Determining the plasma and CSF ?-EP is mea-(ningful) in early diagnosis of CNS infection,and determining the serum CRP at the same time may be helpful in differentiating septic and inseptic infection.

OBJECTIVETo construct the Escherichia coli (E. coli) prokaryotic expression system pET9aHPV11L2E7, purify the fusion protein L2E7 and study the immunnogenicity of the protein.

METHODSThe HPV11 L2, E7 coding region was amplified from condyloma acuminata tissue specimen by PCR. The recombinant plazmid pET9aHPV11L2E7 was established and sequenced. Fusion protein L2E7 (553 amino acids) was expressed in host strain BL21 (DE3plus) by IPTG inducing and identified by using SDS-PAGE and Western blotting. Then L2E7 protein purified with CM column was inoculated to Balb/c mice and its cell-mediated and humoral immunnogenicity was assessed by IFN-gamma enzyme-linked immunospot (ELISPOT) and enzyme-linked immunosorbent assay (ELISA).

RESULTSThe E. coli prokaryotic expression system pET9aHPV11L2E7 was established and the purified fusion protein L2E7 was obtained successfully. The mice in vivo experiment indicated that the purified protein L2E7 could induce HPV11E7 specific cell-mediated immune responses and high level HPV L2E7 antibody was detected in serum.

CONCLUSIONThe purified fusion protein L2E7 could induce specific cell-mediated and humoral immune responses. It can be used as a candidate of genital wart immune therapeutic vaccine.

Animals ; Antibodies, Viral ; blood ; Capsid Proteins ; genetics ; immunology ; isolation & purification ; Escherichia coli ; genetics ; metabolism ; Female ; Gene Expression ; Human papillomavirus 11 ; genetics ; immunology ; Humans ; Mice ; Mice, Inbred BALB C ; Oncogene Proteins, Viral ; genetics ; immunology ; isolation & purification ; Papillomavirus Infections ; immunology ; virology ; Papillomavirus Vaccines ; genetics ; immunology ; isolation & purification ; Recombinant Fusion Proteins ; genetics ; immunology ; isolation & purification

Objective To explore the effect of acupuncture-rehabilitation therapy on apoptosis and the expression of X-linked inhibitor of apoptosis protein (XIAP) and cleaved-caspase-9 in ischemic penumbra of rats with cerebral ischemia. Methods A total of 180 male Sprague-Dawley rats were randomly divided into sham operation group,model group,acupuncture group,rehabilitation group and acupunc-ture-rehabilitation therapy group.Each group was divided into three days,seven days and 14 days subgroups(n=12).The model was estab-lished with the modified Longa suture method. The model group and the sham operation group received no treatment. The acupuncture group received cluster needling of scalp acupuncture,the rehabilitation group received treadmill training,and the acupuncture-rehabilitation therapy group received both acupuncture and treadmill training.On the third,seventh and 14th days after modeling,the neurological func-tion was assessed with modified Neurological Severity Score(mNSS).The apoptosis was observed by TUNEL staining and the expression of XIAP and cleaved-caspase-9 protein was determined by Western blotting in cerebral ischemic penumbra,respectively.Results Compared with the model group,the mNSS scores decreased,the rate of apoptosis decreased,the expression of XIAP protein increased,and the expres-sion of cleaved-caspase-9 decreased in each treatment group at each time point(P<0.05).Compared with the other two treatment groups,the rate of apoptosis further decreased at each time points(P<0.05),the expression of XIAP protein further increased at each time points(P<0.05),the mNSS score further decreased(P<0.05)and the expression of cleaved-caspase-9 protein further decreased(P<0.05)seven days and 14 days after modeling in the acupuncture-rehabilitation therapy group.Conclusion Acupuncture and rehabilitation therapy could re-duce the neurological deficit in rats with cerebral ischemia,which was superior to the simple acupuncture treatment and rehabilitation train-ing.The mechanism may be related to th reduction of apoptosis by promoting XIAP protein expression and inhibiting caspase-9 protein acti-vation.

OBJECTIVETo study the expression of annexin II (AnnII) and the fibrinolytic activity in NB4 cells and their alterations in the presence of arsenic trioxide (ATO) and daunorubicin (DNR).

METHODSLeukemia cell line NB4 was treated with ATO or DNR for 24 ∼ 72 h. Cell surface expression of AnnII and its mRNA were analysed by flow cytometry and real time PCR, respectively, the fibrinolytic activity by chromogenic assay.

RESULTSCompared with other acute leukemia cell lines, the expression of AnnII on untreated NB4 cells was relatively higher. The AnnII positive cell rates on NB4, HL-60, K562, and A3 cells were (94.5 ± 1.6)%, (40.1 ± 2.1)%, (36.3 ± 1.5)% and (11.8 ± 2.5)%, respectively. The fibrinolytic activity of NB4 cells was the greatest with a A value of 0.68 ± 0.02. The fibrinolytic activity of NB4 cells was obviously decreased by ATO, DNR or monoclonal antibody against AnnII, being decreased by 60.4%, 35.8% and 26.0% of the pretreatment level, respectively. The expressions of AnnII and its mRNA in NB4 cells were decreased dramatically after ATO and DNR treated for 48 h. Annexin II positive cells rate were (55.46 ± 4.72)% and (27.00 ± 6.18)%, respectively.

CONCLUSIONNB4 cells have strong ability to enhance the catalytic efficiency of the t-PA-dependent plasminogen activation and AnnII on the cell membrane contributes to this activity. Its high fibrinolytic activity can be corrected by ATO and DNR through down-regulating AnnII.

Annexin A2 ; Apoptosis ; Daunorubicin ; HL-60 Cells ; Humans ; Leukemia ; metabolism

To investigate the role of CD4+, CD25+, and CD127low regulatory T cells (Tregs) in multiple myeloma (MM). Methods:Levels of CD4+T cells and Tregs, as well as expression of CTLA-4 and apoptosis-related proteins, such as CD95, bcl-2, and Caspase3 of Tregs in peripheral blood of 30 patients with newly diagnosed cases, 27 patients under of complete remission (CR) from multiple myeloma patients, and 25 healthy adults were analyzed by flow cytometry. Results:The percentage of CD4+T cells in the untreated group was significantly lower than that of the control group (P<0.05). The percentage of Tregs in CD4+T cells in the untreated group was significantly higher than that of the CR group and control group (P<0.05), which in ISSⅢpatients of the untreated group was significantly higher than that in I/II(P<0.05). No significant difference of CD95 expression in Tregs was observed among the three groups. The expression of CTLA-4 in Tregs from the untreated group was significantly higher than that of the CR group (P<0.05) and control group (P<0.01), and so was in CR group than this in controls (P<0.05). The expression of bcl-2 in Tregs in the untreated group was significantly higher than that of the CR group (P<0.05) and control group (P<0.01), and so was in CR group than this in controls(P<0.05). The expression of Caspase3 in Tregs from the untreated group and CR group were all significantly lower than that of the control group (P<0.05). The percentage of Tregs in CD4+T cells in the untreated group was positively correlated with the proportion of bone marrow plasma cells (P<0.05). The percentage of Tregs in CD4+T cells from 15 MM patients who received bortezamib and dexamethasone (VD) chemotherapy was negatively correlated to the ratio of plasma cell reduction after the first VD chemotherapy (r=0.735, P<0.01). Conclusion:The level of Tregs in the peripheral blood of MM patients was positively correlated with tumor burden and progression of disease, but was negatively correlated with curative effect. The increased level of Tregs was associated with their strengthened anti-apoptosis function.

OBJECTIVETo establish multiple short tandem repeat (STR) amplification by fluorescence labeling polymerase chain reaction (PCR) combined with capillary electrophoresis for quantitative determination of chimerism, and to evaluate the status of engraftment and predict the outcome of allogeneic hematopoietic stem cell transplantation (allo-HSCT).

METHODSThirty-one patients received bone marrow transplantation (BMT) or nonmyeloablative allogeneic stem cell transplantation (NST) were evaluated. Peripheral blood and bone marrow were co-llected before and after transplantation in different period. Nine different STR markers were co-amplified in a single reaction by using a commercial AmpF/STR Profiler Plus PCR amplification kit. Separation of the PCR products and fluorescence detection were performed by ABI prism 310 Genetic Analyzer with capillary electrophoresis. The Genescan and Genotype software were used for size calling and quantification of peak areas. The formula to calculate donor chimerism values was based on the different allelic distribution type between donor and recipient.

RESULTS48.4% of the patients received sex-matched transplantation and the quantification of donor chimerism could only be performed by STR-PCR method. Comparison of values obtained by FISH analysis with that by STR-PCR in patients transplanted from sex-mismatched donors showed an excellent correlation. The median number of informative alleles was 6.7 (range 2 - 10). The donor's alleles appeared in all the patients on day 7 post-transplant. The median values of donor chimerism in BMT group were inferior to that in NST group on day 7, day 14 and 1 month post-transplant. However the difference disappeared in the midterm or later period of transplant. On day 21, all of the 31 patients had stable engraftment and the percentage of donor chimerism was more than 92%. Median follow-up was 17 (3.5 - 29.0) months after transplantation. Twenty-six of 31 patients had durable engraftment and donor chimerism ratio was more than 90%. So for all of them survived leukemia-freely. Four of the 31 patients had unstable mixed chimerism and relapsed within 6 months post allo-HSCT. Another patient with unstable mixed chimerism appeared graft rejection. Decreasing values of donor chimerism were detected prior to the occurrence of graft rejection and disease relapse. The incidence of GVHD was much higher in the group of full donor chimerism.

CONCLUSIONSequential and quantitative monitoring of STR is a valuable tool for studying engraftment dynamics, graft rejection, and relapse and for predicting GVHD. Furthermore it can provide a basis for early intervention of clinical treatment.

Adolescent ; Adult ; Child ; Electrophoresis, Capillary ; Female ; Graft Rejection ; Hematopoietic Stem Cell Transplantation ; Humans ; In Situ Hybridization, Fluorescence ; Male ; Middle Aged ; Polymerase Chain Reaction ; Recurrence ; Tandem Repeat Sequences ; Transplantation Chimera ; Transplantation, Homologous

OBJECTIVETo evaluate the effects of egg and milk supplementation on growth and development and body composition among children in poor rural area in Tianyang County of Guangxi province.

METHODSTotal four schools were randomly selected from four towns in Tianyang County of Guangxi province as intervention group in April, 2013. The intervention measures included that these students were given salty egg (net weight: 50 g) and ultra-high-temperature-sterilization school milk (net weight: 200 g) every school day and these schools were equipped with standard kitchens. Another four schools of familiar socio-economic level, teaching quality and size from the same town were randomly chosen as control group and none of the intervention measures were implemented. About 25 students were randomly selected and stratified by grades from grade one to grade five. The height, weight, and body composition of all students were measured in April, 2013 and one year after the intervention. A total of 978 students were measured at baseline from age 6 to 13, 552 students as intervention group and 426 as control group. t-test was used to compare the differences between groups and multivariate unconditional logistic regression was used to analyze the factors of malnutrition.

RESULTSAfter one year intervention, 892 students were measured randomly, with 515 students in intervention group and 377 in control one. The average weight of boys in intervention group increased (3.6 ± 1.7) kg compared with baseline. It was significantly higher than that of control group ((2.9 ± 1.5) kg) (t = 4.40, P < 0.001). The boy's lean body mass of intervention group increased (2.6 ± 1.4) kg, higher than the control group ((2.0 ± 1.2) kg) (t = 3.95, P < 0.001). The decrease of malnutrition rate of intervention schools (11.8%) was significantly higher than that of the control schools (4.7%, χ² = 16.90, P < 0.001), and the odds ratio was 0.37 (95% CI: 0.23-0.59). The risk difference of overweight and obesity was not statistically significant between the two groups (OR = 1.68, 95% CI: 0.57-4.94).

CONCLUSIONAfter supplementing milk and egg, the nutritional status of the poor rural pupils was improved.

Animals ; Body Composition ; Body Weight ; Child ; Child Development ; China ; Diet ; Eggs ; Humans ; Male ; Milk ; Nutritional Status ; Obesity ; Overweight ; Poverty Areas ; Rural Population ; Schools ; Students

OBJECTIVETo investigate the protective effects of estrogen on the mitochondria in human umbilical vascular endothelial cells (HUVECs).

METHODSHUVECs were exposed to H2O2 at 250 micromol/L for 4 h with or without pretreatment with 17-estradiol (E2) and ICI182780. Complex IV activity of the cells was measured with chromometry, and 2, 7-dichlorofluorescein diacetate (DCFH-DA) was used to determine intracellular reactive oxygen species (ROS). Intracellular adenosine triphosphate (ATP) level was quantified with a luciferin- and luciferase-based assay.

RESULTSCompared to the blank control group, H2O2 caused a decrease in complex IV activity, intracellular ATP level, and the cell viability, but elevated intracellular ROS. E2 pretreatment of cells significantly attenuated these effects of H2O2 exposure. ICI182780 administered prior to E2 pretreatment antagonized the protective effects of E2 against H2O2 exposure.

CONCLUSIONE2 offers mitochondrial protective effects on HUVECs, which is mediated by the estrogen receptors.

Cells, Cultured ; Cytoprotection ; drug effects ; Electron Transport Complex IV ; metabolism ; Endothelial Cells ; cytology ; drug effects ; metabolism ; Estrogens ; pharmacology ; Female ; Humans ; Hydrogen Peroxide ; pharmacology ; Mitochondria ; drug effects ; metabolism ; Oxidative Stress ; drug effects ; Pregnancy ; Reactive Oxygen Species ; metabolism ; Umbilical Veins ; cytology