Objective To investigate the role of IgG determination in diagnosis of early renal damage in children with Henoch-schonlein purpura(HSP). Methods Urine IgG was determined in 188 children with HSP (HSP group), 99 patients with Henoch-schonlein purpura nephritis (HSPN group) and 26 healthy controls (healthy control group). Simultaneously, qualitative detection of urine protein was carried out in HSP group. Results The abnormality rate of urine IgG was significantly higher than that of urine protein in HSP group (P<0. 05). The level of urine IgG was significantly higher in HSP group than that in healthy control group (P<0. 05), while the level of urine IgG in group than that in HSP group (P<0. 05). Conclusion Urine IgG determination is simple and nonin-vasive, which if high clinical value in diagnosis of early renal damage in children with Henoch-schonlein purpura(HSP).

[Abstract ] Objective Globular C1q receptor (gC1qR), a highly acidic receptor protein, is expressed in almost all mamma-lian cells in addition to exoerythrocytic, which can mediate a variety of biological responses.The study aimed to explore gC1qR expres-sion in cervical cancer and itd effects on the biological behaviors of human cervical cancer cells. Methods Retrospective analysis was made on 100 cervical tissue samples of patients in Nanjing Maternal and Child Health Hospital from August 2014 to April 2015, in-cluding 50 cervicitis tissues and 50 cervical cancer tissues.Immunohistochemical SP method was applied in the research of cervical cancer cell line C33a to detect the expression and the location of gC1qR in cervical tissues.Real-time PCR and Western blot analysis were respectively applied to detect the levels of gC1qR mRNA and gC1qR protein expression.Besides, the abilities of C33a cells mi-gration, invasion and apoptosis were respectively assessed by in vitro cell wound healing experiment, transwell assay and flow cytome-try. Results The expression of gC1qR gene was dramatically decreased in the group of cervical cancer tissues when compared with chronic cervictis group (2.18 ±0.37 vs 7.23 ±0.69, 0.27 ±0.09 vs 0.74 ±0.02, P<0.001).gC1qR overexpression could result in sig-nificant up-regulation of cervical celluer apoptosis([22.89 ±1.46]%vs [12.98 ±0.57]%) and down-regulation in migration ([42.60 ± 3.29]%vs [141.83 ±4.71]%) and invasion([26.20 ±2.89]%vs [67.13 ±0.95]%).Typical apoptosis was also observed in cervi-
cal cells by transmission electron microscope. Conclusion gC1qR expression might play an important role in inhibiting the invasion and migration of cervical cancer cells and inducing the apoptosis of cervical carcinoma cells, which provides new clues and potential targets for the treatment of cervical cancer in further research.

Nobel Prize 2009 in Physiology or Medicine is awarded to three American scientists, Elizabeth H.Blackburn, Carol W.Greider and Jack W.Szostak, for the discovery of"how the chromosomes are protected by telomeres and the enzyme telomerase".Telomere is the specific structure at the ends of the chromosomes and protects it from fusion and degradation.Telomerase synthesizes telomere DNA to maintain the telomere length.Studies suggest that telomere length and telomerase activity is directly associated with cell life and the genesis of many diseases.With the progress of study, how to control the telomere length and telomerase activity is helpful to shed light on the studies in"cancer, inherited diseases and senescence", and will stimulate the development of potential new therapies.

Objective To establish the triplex Taqman probes real-time RT-PCR method for simultaneously detecting of EV71,CA16 and EV.Methods Retrospective study.Specific primers and probes were designed based on conserved regions of EV71,CA16 and EV.The sensitivity,specificity and reproducibility were assessed by the optimized reaction system.A total of 176 throat swabs as the experimental group were collected from children with suspected hand foot mouth disease (HFMD),who admitted from April 2012 to July 2012 in Nanjing Children's Hospital affiliated to Nanjing Medical University.During this time,10 cases of healthy children,10 cases of outpatients with flu-like symptoms and 90 cases of inpatients in pneumology department of our hospital were recruited as control group,whose throat swabs were also collected.All of 286 samples were tested by the triplex Taqman probes real-time RT-PCR for simultaneously detecting EV71,CA16 and EV.SPSS13.0 was used to analyze the results.Results The sensitivities of the triplex Taqman probes real-time RT-PCR was 1.0 × 103 copies per milliliter for EV71,CA16 and EV.It showed 100％ specificity for 9 enterovirus and 3 non-enterovirus.Analysis with 1.0 × 103-1.0 × 105 copies per milliliter constructed plasmids demonstrated high reproducibility with coefficient of variation of 0.44％-1.04％ for EV71,0.38％-0.73％ for CA16,and 0.46％-0.90％ for EV.More over 176 samples collected from children with suspected HFMD were detected by triplex Taqman probes real-time RTPCR and real-time RT-PCR.The results showed 97.2％ (171/176)agreement and 0.94 Kappa value with high concordance.Conclusions The triplex Taqman probes real-time RT-PCR detecting EV71,CA16 and EV simultaneously has been established successfully.The assay,with high sensitivity and specificity,provide good basis for the rapid clinical diagnosis of EV71,CA16 and EV and open up broad prospects for clinical and relevant researches.

Objective To explore the relationship between peripheral Th17 cell number and chronic gastritis in Helicobacter pylori(H.pylori)-infected children.Methods Children were diagnosed as chronic gastritis by endoscopy.The degree and activity of inflammation were graded by histopathology examinations.The patients with both 13C urea breath test and urease test positive were diagnosed as H.pylori infection.The peripheral Th17 cell number was measured by flow cytometry and expressed as a ratio to total T cell.Results The Th17 cell number in HP group (chronic gastritis with H.pylori infection,n =33),non-HP group (chronic gastritis without H.pylori infection,n =24) and normal controls (n =15) were (1.55 ±0.30)％,(1.06 ±0.33)％,and (1.04 ±0.35)％,respectively.HP group included a statistically higher Th17 cell number than the other groups (all P ＜ 0.05),while no obvious difference was found between non-HP group and controls (P ＞ 0.05).According to the degree of inflammation,the chronic gastritis with H.pylori infection was categorized into non-apparent (n =10),mild (n =8),moderate (n =9) and severe (n =6) subgroups.The Th17 cell number in each subgroup was (1.64 ± 0.21)％ (non-apparent),(1.61 ± 0.23)％(mild),(1.25 ± 0.29) ％ (moderate) and (1.75 ± 0.20) ％ (severe),respectively.The moderate group had a lowest Th17 cell number among 4 groups (P ＜ 0.05).And significant differences did not exit in the other 3 groups (P ＞ 0.05).The HP group patients with different inflammatory activity had a Th17 cell number of (1.23 ±0.25)％ in nonapparent (n=15),(1.53 ±0.15)％ in mild (n=6),(1.55 ±0.32)％ in moderate (n=6) and (1.71 ±0.35)％ in severe (n =6) subgroup,respectively.However,there were no significant differences among 4 subgroups (P ＞ 0.05).Conclusions In the progress of chronic gastritis with H.pylori infection,Th17 cells may play a role as a double-edged sword by protecting and fighting against H.pylori infection and immunopathologic insults.This would provide more insights into the treatment of H.pylori infection.

Objective To explore the role of long noncoding RNA ( lncRNA) CTD-2012K14. 6 in the development of gestational diabetes mellitus (GDM) related macrosomia. Methods The quantitative real-time PCR ( qRT-PCR) was performed to measure the expression of CTD-2012K14.6 in placentas of women with or without GDM, and the quantity of CTD-2012K14. 6 expression and its association with fetal weights were analyzed; Bioinformatic analysis was performed to predict the downstream molecules. CTD-2012K14. 6 over-expressing lentiviral and siRNA was constructed in human trophoblastic cell line HTR-8/SVneo cells, qRT-PCR and Western blot (WB) were used to invest its effect in modulating the expression of downstream molecules. Results The expression of CTD-2012K14.6 in GDM placentas was significantly higher than that in normal controls (1.70 ± 0.63 vs 1.00 ± 0.56,t=3.68,P＜0.01), and positively correlated with fetal weight (r=0.8501, P＜0.01); on-line analysis showed that CTD-2012K14.6 was located at chr16:67,549,214-67,563,958, which was located in the intron of CCCTC-binding factor( CTCF); Up-regulating CTD-2012K14.6 could significantly reduce the expression of CTCF mRNA and protein, and increase the expression of insulin-like growth factor-Ⅱ( IGF-Ⅱ) mRNA and protein, while down-regulating CTD-2012K14.6 could significantly increase the expression of CTCF mRNA and protein, and reduce the expression of IGF-ⅡmRNA and protein. Conclusion The CTD-2012K14. 6 may play an important role in the pathogenesis of GDM related macrosomia by upregulating the expression of CTCF and IGF-Ⅱ.

AIM: To study the effect of the overexpression of p16 on an anion exchange function of band 3 in HeLa cells. METHODS: The expression of p16 and band 3 in HeLa cells was detected by immunohistochemistry (IHC). The p16 cDNA was subcloned to plasmids pEGFP-C1 by PCR and identified by restriction enzyme digestion and sequencing, and then, the recombinant pEGFP-C1-p16 plasmids were transiently transfected into HeLa cells. The expression of fusion protein in HeLa cells was detected by fluorescence microscope. 6-methoxy-N-(3-sulfopropyl)-quinolinium(SPQ)fluorescent probes were used to detect the anion exchange function of band 3. RESULTS: P16 and band 3 were expressed in HeLa cells. The amplificated p16 cDNA sequence was the same as the report sequence. The transfective efficacy of pEGFP-C1-p16 was above 60%. The anion exchange function increased after the transfection of pEGFP-C1-p16 plasmids. CONCLUSION: p16 facilitates the anion exchange function of band 3 in HeLa cells.

Objective To analyze the positive report time,distribution and antimicrobial resistance of pathogens isolated from blood culture at a hospital,so as to provide laboratory basis for prevention,contro1 ,and rational antimicrobialuse for bloodstream infection.Methods From January 2013 to January 2015,blood culture specimens of outpatients and inpatients were performed bacterial identification and antimicrobial susceptibility testing, antimicrobial resistance was analyzed.Results A total of 1 973 blood culture specimens were sent by clinical depart-ments,219 (11 .10%)of which were isolated pathogens.Most positive blood culture specimens were from depart-ment of paediatrics (n = 199 ).Isolation rates of gram-negative bacteria,gram-positive bacteria,and fungi were 44.34% (n=98),50.23% (n=111),and 5.43% (n=12)respectively;the main pathogens was coagulase-negative staphylococcus (n=53,23.98%),followed by Escherichia coli (n=39,17.65%),Staphylococcus aureus (n=23, 10.41 %),Klebsiella pneumoniae (n =15,6.79%),and Pseudomonas aeruginosa (n =13,5.88%),the average positive blood culture report time of top five pathogens was 1 -2 days.The detection rates of extended-spectrumβ-lactamase-producing Escherichia coli and Klebsiella pneumoniae accounted for 53.85% and 53.33% respectively, susceptibility of gram-negative bacilli to carbapenems was relatively high(76.92% - 100%);methicillin-resistant isolates accounted for 39.13% among Staphylococcus aureus and 64.15% among coagulase-negative staphylococ-cus,vancomycin-resistant and teicoplanin-resistant strains were not found;resistant rate of Candida glabrata to 5-fluorocytosine was 14.29%,but was susceptible to amphotericin B.Conclusion The major pathogens isolated blood culture are gram-positive bacteria,in order to reduce the emergence of drug-resistant strains,clinicians should choose antimicrobial agents according to blood culture results and antimicrobial susceptibility testing results.

Objective To investigate the coverage of a recombinant protein vaccine based on pneumococcal surface protein A (PspA) from both family 1 and family 2.Methods One hundred and fifty-nine Streptococcus pneumoniae strains, including 47 invasive strains, were isolated from children in Nanjing Children′s Hospital.Cell lysates were prepared and reacted with three antibodies recognizing PspA -RX1, PspA-3296 and PspA-5668 for PspA typing by ELISA .Results Among 47 invasive isolates of 9 different serotypes, 10.7%were PspA family 1 and 89.3%were PspA family 2.Among all of 159 clinical isolates, 10.1% were identified as PspA family 1, 88.0%were family 2, while 1.9%of strains could not be typed by ELISA and PCR assays .None of strains belonged to PspA family 3.Conclusion The recombinant pro-tein vaccine based on PspA from both family 1 and family 2 has a broad coverage among clinical isolates and is potentially protective against both invasive and non-invasive pneumococcal diseases .

Vitamin is one of the essential nutrients of human body.Monitoring the level of the vitamin receives more and more attention clinically.Common ways of vitamin detection include microbiological method, fluorescence analysis, immunological method, high performance liquid chromatography,liquid chromatography-mass spectrometry,electrochemical method and so on.In addition, there′re still many disputes on the reference interval of vitamins,the medical decision level and the specific reference intervals for different groups.Therefore,establishing a standardized method of vitamin detection is an urgent problem to be solved.