Rapid diagnosis is an important link in the prevention and control of infectious diseases.Point-of-care testing(POCT)is portable, fast, easy to operate, intelligent and sensitive, which has been widely used in the detection of pathogenic microorganisms of infectious diseases, host biomarkers, microbial drug sensitivity in recent years.It is of great significance for the monitoring and management of disease epidemiology and rational use of antibiotics.This review summarized the application of POCT in the diagnosis and treatment of pediatric infectious diseases.

Objective To investigate clinical significance of soluble CD30/CD30L and CD40/CD40L system imbalance in ovarian serous tumors.Methods 40 patients of serous cystadenoma and 30 patients of serous cystadenocarcinoma were selected,and 40 age-and weight-matched healthy women were also recruited as the control group.Peripheral venous blood (3 ml) of the healthy control and patients with ovarian serous tumors before surgery and 7 days after surgery were collected.After separation of serum,ELISA was used to detect levels of sCD30,sCD30L,sCD40 and sCD40L.Results Compared to the control group,levels of sCD30,sCD30L,sCD40 and sCD40L in both serous cystadenoma and serous cystadenocarcinoma groups were significantly in creased (P＜0.05).And in those serous cystadenocarcinoma group,levels of such soluble proteins were much higher than in serous cystadenoma group (P＜0.05).7 days after surgery,levels of such soluble proteins were significantly decreased in both serous cystadenoma and serous cystadenocarcinoma groups (P＜0.05).Conclusion Detection of serum sCD30/sCD30L and sCD40/sCD40L is possible to have a certain guiding significance to early diagnosis of ovarian tumors and the prognosis of patients.

Objective:To explore side population(SP) cells in human epithelial cells and to observe the expression of the universal stem cell marker ABCG2,epidermal stem cell markers ?6 integrin and ?1 integrin on SP cells.Methods:Epithelial cells were obtained by digesting human skins with Dispase II and Trypsin and stained with Hoechst33342 and PI.The SP cells were analyzed and sorted by the fluorescence-activated cell sorter.Then the expression of ABCG2,?6 integrin and ?1 integrin was analyzed by flow cytometry.Results:SP cells accounted for 0.2%-0.3% of total human epithelial cells.Only a small part of cells expressed ABCG2,integrin ?6 and integrin ?1 of both SP cells and total human epithelial cells detected by flow cytometry.The difference of positive rates between SP cells and total human epithelial cells was not significant.Conclusion:SP cells accounted for 0.2%-0.3% of total human epithelial cells.The difference of positive rates of stem cells marker ABCG2,integrin ?6 and integrin ?1 between SP cells and total human epithelial cells was not significant.

Objective To investigate the characteristics and distribution of class teaching skill points in state high quality course pathology using Flanders interaction analysis system to analyze the teaching video of pathology course.Methods Teaching videos of state high quality course (pathology course) between 2006 and 2012 were analyzed by Flanders interaction analysis system.Frequencies of using teaching skill,presentation skill and interaction skill were statistically analyzed.Results The study found that frequencies of using teaching skill points in pathology course were decreased in the following order:presentation skill (53.94％),interaction skill (26.64％) and teaching skill (19.42％).Sensory focusing was most commonly used in presentation skill.Monitoring and expressing concern were most commonly used in interaction skill.Gestures and cases were most commonly used in teaching skill.Conclusions Teachers in pathology teaching should correctly handle the relationship among the presentation,interaction and teaching skills according to the teaching objectives and teaching content.

Objective To explore the characteristics and clinical significance of CD+4 CD+25 Regulatory T lymphocytes and T cell subsets in peripheral blood and malignant pleural effusion from lung cancer patients. Methods Flow cytometry was used to detect the percentage of CD+4 CD+25 regulatory T cells and T cell subsets in peripheral blood from 68 lung cancer patients and 56 healthy persons, and in pleural effusion from 32 lung cancer patients with malignant effusion. Results T lymphocyte subsets in peripheral blood of lung cancer patients in different periods were expressed differently. The percentage of CD+4 CD+25 regulatory T cells in peripheral blood were (19.52±3.32)%, (27.28±8.26)% and (32.31±15.60)% in Ⅰ+Ⅱ, Ⅲ and Ⅳ period lung cancer patients, respectively, and were higher than that of healthy volunteers (11.12±3.32) % (t =31.0040, -7.9688, -4.9770, P ＜0.05). In the lung cancer patients with malignant effusion, the percentage of CD+4 CD+25 regulatory T cells in the pleural effusion was higher than that in the peripheral blood [(34.12±18.63) % vs (26.36± 16.25)%, t =21.164, P＜0.05]. In the lung cancer patients with malignant effusion ,the percentages of CD+4 in peripheral blood and pleural effusion were (25.32±13.45) % and (34.68±12.34) %, were lower than that in healthy volunteers (t =7.3104, 4.8818, P＜0.05), the percentages of CD+56 were (8.24±7.38) % and(11.23± 7.65) %, CD+4/CD+8 were (1.02±0.56) % and (1.32±0.82)%, were lower than (18.23±9.23) % and (1.89± 0.32) % in healthy volunteers, respectively, (CD+56: t =-14.7549, -11.7216; CD+4/CD+8: t =-24.78,-4.4564, P＜0.05). Conclusion The relative increase of CD+4 CD+25 Regulatory T cells may be related to immunosuppression and tumor progression in patients with lung cancer. Conclusion The relative increase of CD+4 CD+25 regulatory T cells may be related to immunosuppression and tumor progression in patients with lung cancer.

Multiple kinds of Artificial Skin Substitute are now available. However, except for the Homo Skin Graft there is no Artificial Skin Substitute that can be used as permanent Artificial Skin Substitute. During the past 20 years, more and more scholars around the world have expressed increased interests in the research and development of Artificial Skin Graft that can be utilized as satisfying permanent Artificial Skin Substitute. We conducted our research on the biological evaluation of medical devices of Collagen-Chitosan(C-C) Artificial Skin Substitute according to the National Standard (GB/T16886. 1-1997). The following experiments were conducted: (1)Cytotoxicity, (2)Systemic toxicity(acute toxicity), (3)Haemocompatibility, (4)Sensitization, (5)Intracutaneous reactivity, (6)Pyrogen test, (7)Genotoxicity. The experiment results demonstrate that all biological functional indexes of the Artificial Skin Graft meet the National Standards. Therefore, we conclude that C-C Artificial Skin Graft is characteristic of good biological compatibility. It is non-irritant and has no systemic and cellular toxicity, no genotoxicity, no pyrogen, and no allergen.
Animals ; Chitosan ; toxicity ; Collagen ; toxicity ; Female ; Male ; Materials Testing ; Mice ; Random Allocation ; Skin, Artificial ; adverse effects

OBJECTIVETo develop a recombinant single domain antibody against hTERT, human telomerase catalytic subunit.

METHODSA previously prepared His-tagged hTERT fusion protein was used as the antigen, and the variable regions in heavy chain (VH) of immunized mice were RT-PCR amplified and cloned into the pCANTAB 5E, a phagemid vector. By transfection, the display library of mouse VH was developed. The candidate clones were selected by affinity panning, and soluble VH were obtained after expression in E. coli, HB2151. The resultant single VH antibodies were characterized on their binding potentials by western blotting.

RESULTSAn about 350 bp VH fragment was amplified from spleen cells of mice immunized by His-tagged hTERT and expressed by phage displayed as VH library. The size of the library was 8 x 10(4). After three rounds of affinity panning, 4 independent clones were chosen and consequently expressed as soluble single domain antibodies (Mr = 16 000). In Western blot analysis, the single domain antibody from 2 of 4 clones proved to react with the His-tagged hTERT fusion protein (Mr = 167 000) without dependence of His-tags and also detect the native hTERT (Mr = 127 000) extracted from the human HeLa cancer cell line. DNA sequencing showed both of the single domain antibodies were encoded by the heavy chain variable region of the mouse.

CONCLUSIONSThe single domain antibodies developed were hTERT recognizable and hTERT specific, thus providing a basis for application of recombinant single domain antibody in inhibition of telomerase activity and anticancer therapy.

Amino Acid Sequence ; Animals ; Antibodies, Monoclonal ; genetics ; immunology ; Base Sequence ; Cloning, Molecular ; Complementarity Determining Regions ; genetics ; DNA-Binding Proteins ; HeLa Cells ; Humans ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin Variable Region ; genetics ; Mice ; Molecular Sequence Data ; Sequence Analysis, DNA ; Telomerase ; immunology

Objective To evaluate the effect of cardiopulmonary bypass (CPB) on the expression of hypothalamic nerve growth factor precursor (proNGF) and the influence of hypothalamic proNGF on the sympathetic output of paraventrucular nucleus. Methods Forty-two male SD rats, aged 3-4 months, weighing 350-500 g, were divided into control group, CPB group and ischemia reperfusion (IR) group. At the end of CPB for 110 min, hypothalamus and dorsal root ganglion (DRG) were taken to measure the levels of proNGF mRNA and hypothalamic proNGF protein. Mini pipe was put into bilateral paraventrucular nucleus (PVN) and human recombination proNGF protein was injected into PVN for 7 d before the local field potentials (LFP) of RVLM was recoreded. Human recombination proNGF protein was administrated into lateral ventricle, the prior-administration-LFP of PVN and post-administration-LFP were recorded and compared. At the end of the experiment, hypothalamus was taken to measure the levels of glutamate and gammer amino butyric acid (GABA). Results Hypothalamic proNGF protein in CPB group and IR group was higher than that in the control group (P ＜ 0.05); NGF mRNA of hypothalamus and DRG in CPB and IR group were higher than those of control group (P ＜ 0.05). In PVN and RVLM, after the administration of proNGF protein, the power of delta band significantly decreased and other bands increased (P ＜ 0.05). The hypothalamic GABA level decreased (P ＜ 0.05) with no change of hypothalamic glutamate after proNGF was injected into lateral ventricle. Conclusion CPB increases the expression of proNGF in the hypothalamus contributing to the changes of hypothalamic sympathetic output.

Objective:To evaluate the biosafety of medical injectable carboxymethyl glycosaminoglycan gel.Methods:Ames test, chromosome aberration test in vitro and gene mutation test in vitro were used to detect the genotoxicity of the medical carboxymethyl glycosaminoglycan gel. The gel saline extract (50 ml/kg) was injected slowly through the marginal vein of the ear into Japanese big-eared rabbits. The body temperature was measured and the temperature rise was calculated. The gel saline extract (50 ml/kg) and normal saline (control) were injected intraperitoneally and intravenously into the Kunming mice, respectively. The toxicity response in mice was observed after injection, and bodyweight change was valued. The gel saline extract, normal saline and distilled water were added into the rabbit anti-clotting, to detect the rate of hemolysis.Results:Under active and inactive conditions, the number of spontaneous revertants of the 4 strains of gel saline extract group and gel DMSO extract group did not reach 2 times of that of the corresponding negative control group. The rate of chromosome aberration of the three dose groups were 0. There was no significant increase in the large colony mutation frequency, small colony mutation frequency and total mutation frequency in three dose groups (all P>0.05). After injection of gel saline extract for 24, 48 and 72 h, no toxic reaction was found in each group of mice. With the extension of time after injection, the body weight of mice in the sample group and the control group increased, but the difference was not statistically significant ( P>0.05). After injection of gel saline extract, the temperature rise of 3 Japanese big-eared rabbits were 0.0, 0.3 and 0.2 ℃ respectively. The results of hemolysis test showed that the hemolysis rate of the polycarboxymethyl glucosamine gel was 0.1%. Conclusions:No genetic toxicity changes were found in carboxymethyl glycosaminoglycan to induce gene mutation or chromosome damage in bacteria and cells, and no pyrogenicity, acute systemic toxicity and hemolysis were observed. These results indicate that thecarboxymethyl glycosaminoglycan gel has good biosafety.

OBJECTIVE: To analyze the pathogenic variant of preaxial polydactyly in a Chinese Han pedigree and identify the cause of polydactyly. METHODS: The peripheral blood DNA of the proband and her parents was extracted. The polydactyly-related genes were detected by trio whole exome sequencing, and the suspected pathogenic gene was screened out. Sanger sequencing was applied to other members of the pedigree. RESULTS: The results of gene sequencing showed that the LMBR1 gene had a heterozygous variant of c.423+4909(IVS5)C>T in 6 patients of the pedigree. The same variant was not detected in family members with normal phenotype. Based on the ACMG guidelines, c.423+4909(IVS5)C>T of the LMBR1 gene was predicted to be pathogenic (PM1+PM2+PP1-S(PS)+PP4+PP5). CONCLUSION The heterozygous C>T variant at position 4909 of intron 5 of the LMBR1 gene probably underlies the disease in this pedigree.
China ; Female ; Humans ; Mutation ; Pedigree ; Polydactyly/genetics* ; Thumb ; Whole Exome Sequencing