OBJECTIVETo verify the binding of p53 to p21WAF1/CIP1 gene promoter and detect its binding to hsp90 beta gene promoter in vivo.

METHODSChromatin immunoprecipitation and PCR analysis were used to measure specific gene regulation sequence and Western blot analysis to investigate p53 protein.

RESULTSThe p53 binding sequences on the promoters of p21WAF1/CIP1 and hsp90 beta gene were found in the p53 antibody immunoprecipitated DNA fragments and p53 was detected in the immunoprecipitated samples.

CONCLUSIONSp53 binds to promoters of p21WAF1/CIP1 and hsp90 beta gene in vivo, and regulates the expression of the two genes.

Binding Sites ; Chromatin ; genetics ; isolation & purification ; Cyclin-Dependent Kinase Inhibitor p21 ; Cyclins ; genetics ; metabolism ; Gene Expression Regulation ; HSP90 Heat-Shock Proteins ; genetics ; metabolism ; Humans ; Polymerase Chain Reaction ; methods ; Precipitin Tests ; methods ; Promoter Regions, Genetic ; Protein Binding ; Transcription, Genetic ; Tumor Suppressor Protein p53 ; genetics ; metabolism ; physiology

The bioinformatics software was used to predict the B cell and T cell epitopes of Toxoplasma gondii microneme 16 (TgMIC16) followed by epitopes display on the yeast of Saccharomyces cerevisiae.B and T cell epitopes of TgMIC16 were predicted by DNAStar and IEDB,and software of SYFPEITHI and BIMAS,respectively.According to the predicted results,the antigenic epitope region was expressed with pCTCON2/EBY100 display system.The expression protein was detected by indirect immunofluorescence (IFA) and flow cytometry.Results showed that there were potential B/T cell epitopes in TgMIC16 and concentrated in the aa343-625 region.The epitope region was successfully displayed on the surface of yeast cells,and the optimal induction time was 24 hours.The study would lay the foundation for the development and efficacy evaluation of the yeast carrier vaccine in the further research.

OBJECTIVETo investigate the clinical results of the treatment of severe infantile hemangioma with high-dose propranolol in Chinese.

METHODS56 cases with severe infantile hemangioma were treated with propranolol. Clinical evaluation, electrocardiography, and experimental examination of liver function and heart function were performed before treatment. The daily dose of propranolol was increased from 1 mg/kg at the first day to 1.5 mg/kg at the second day, and to 2 mg/kg at the third day. The propranolol was given twice a day. The treatment was lasted for six months. The patients were visited every month.

RESULTSThe lesion color was changed after 2-4 days of treatment in all the cases. All the lesions were dramatically improved after one month of treatment. The ulceration were healed, except one case. Until now, complete regression was achieved in 10 cases and marked improvement in 46 cases. Side effects were happened in 3 cases, including one case of abnormal liver function, one case of CK-MB increase and one case of continuous increase of CK-MB, LDH, ALT, GGT.

CONCLUSIONSHigh-dose Propranolol is very effective in the treatment of infantile hemangioma with minor side effects and short disease period. It might he used as the first-line treatment for infantile hemangioma.

China ; Female ; Hemangioma ; drug therapy ; Humans ; Infant ; Male ; Propranolol ; administration & dosage ; therapeutic use ; Treatment Outcome

OBJECTIVETo investigate the prevalence of hearing impairment and ear diseases in old people and provide scientific data for drawing up the prevention and treatment strategies.

METHODSUsing the probability proportion to size (PPS) method, 1261 people over 60 years were investigated in 40 clusters in Jiangsu Province with the WHO protocol.

RESULTSThe prevalence of hearing impairment was 58.1% (the standardized rate: 59.5% in the whole country, 60.9% in Jiangsu province). Degrees of hearing impairment were mild (33.1%), moderate (17.8%), severe (5.9%) and profound (1.3%). The prevalence of hearing disability was 25.0% (the standardized rate: 26.6% in the whole country, 28.1% in Jiangsu province). There were significant difference of the prevalence between male and female, as well as urban and rural, and different ages. The prevalence of the ear diseases was auricle malformation (0.2%), wax (1.7%), otitis externa (0.1%), fungi (0.5%), serous otitis media (1.2%), chronic suppurative otitis media (1.6%), dry perforation of tympanic membrance (2.3%). The causes of hearing impairment were ear diseases (2.9%), non-infectious condition (92.6%), genetic condition (0.3%) and undetermined causes (4.2%). Of which, 31.1% of persons needed hearing aids while 2.3% of persons needed medicine treatment, but 0.9% of persons needed non-urgent surgery and 1.0% of persons needed other treatment.

CONCLUSIONSThe prevalence of hearing impairment and disability in the old rised obviously than the last investigation in 1987. It was a heavy burden for social development in China. The government and the whole society should take more concern about the problem. The scientific strategies of prevention and treatment were urgently needed and implemented.

Aged ; Aged, 80 and over ; Audiometry, Pure-Tone ; China ; epidemiology ; Ear Diseases ; epidemiology ; Female ; Hearing Loss ; epidemiology ; Humans ; Male ; Middle Aged ; Prevalence

Malaria remains a serious public health problem in Shandong Province, China; therefore, it is important to explore the characteristics of the current malaria prevalence situation in the province. In this study, data of malaria cases reported in Shandong during 2012-2014 were analyzed, and Plasmodium species were confirmed by smear microscopy and nested-PCR. A total of 374 malaria cases were reported, 80.8% of which were reported from 6 prefectures. Of all cases, P. falciparum was dominant (81.3%), followed by P. vivax (11.8%); P. ovale and P. malariae together accounted for 6.4% of cases. Notably, for the first time since 2012, no indigenous case had been reported in Shandong Province, a situation that continued through 2014. Total 95.2% of cases were imported from Africa. The ratio of male/female was 92.5:1, and 96.8% of cases occurred in people 20-54 years of age. Farmers or laborers represented 77.5% of cases. No significant trends of monthly pattern were found in the reported cases. All patients were in good condition after treatment, except for 3 who died. These results indicate that imported malaria has increased significantly since 2012 in Shandong Province, especially for P. falciparum, and there is an emergence of species diversity.
Africa ; China* ; Farmers ; Humans ; Malaria* ; Microscopy ; Plasmodium falciparum ; Plasmodium malariae ; Plasmodium ovale ; Plasmodium vivax ; Plasmodium* ; Prevalence ; Public Health

Objective To construct pEGFP-N1-HBsAg-ROP2 recombinant expression plasmid and transfect HEK293T cells for expression,and pay a way for Toxoplasma gondii nucleic acid vaccine development. Methods According to the HBsAg gene sequence and pcDNA3-p30-ROP2 recombinant plasmid restriction sites,the HBsAg gene was amplified by PCR.The HB-sAg gene was cloned into the pcDNA3-p30-ROP2 and instead of p30 gene.The HBsAg-ROP2 fragment was amplified by PCR and digested with HindⅢand KpnⅠto clone into the pEGFP-N1 eukaryotic expression vector and construct the recombinant pEGFP-N1-HBsAg-ROP2.The expression vector was transfected into HEK293T cells based on the identification of PCR amplifi-cation,restriction endonucleases and sequencing.Results The PCR product of HBsAg was about 700 bp,which was consis-tent with the theoretical value.Two bands of about 5.4 kb and 1.9 kb were obtained after double enzyme digestion with pcDNA3-HBsAg-ROP2 recombinant plasmid.The recombinant plasmid pEGFP-N1-HBsAg-ROP2 was double-digested to generate an empty vector fragment of about 4.7 kb and a band of about 1.9 kb of HBsAg-ROP2 fragment.The results of sequencing showed that the sequence was 99.84% identical with the published sequence in GenBank.The target plasmid was successfully transfect-ed into HEK293T cells,and the expression was correct,the protein concentration was 3.08 mg/ml.Conclusion The recombi-nant plasmid pEGFP-N1-HBsAg-ROP2 is successfully constructed and expressed efficiently.

A large number of protease inhibitors have been found from leeches, which are essential in various physiological and biological processes. In the curret study, a novel elastase inhibitor was purified and characterized from the leech of Hirudinaria manillensis, which was named HMEI-A. Primary structure analysis showed that HMEI-A belonged to a new family of proteins. HMEI-A exerted inhibitory effects on elastase and showed potent abilities to inhibit elastase with an inhibition constant (K
Amino Acid Sequence ; Animals ; Leeches/chemistry* ; Pancreatic Elastase/antagonists & inhibitors* ; Protease Inhibitors/pharmacology* ; Proteins

ObjectiveTo investigate the drug-resistant gene polymorphisms in Plasmodium falciparum imported from Equatorial Guinea to Shandong Province. MethodsFrom 2015 to 2016, blood samples were collected from imported P. falciparum malaria patients returning from Equatorial Guinea to Shandong Province, and genome DNA of the malaria parasite was extracted. The drug-resistant Pfcrt, Pfmdr1, Pfdhfr, Pfdhps, and K13 genes of P. falciparum were amplified using a PCR assay, followed by DNA sequencing, and the sequences were aligned. Results The target fragments of all 5 drug-resistant genes of P. falciparum were successfully amplified and sequenced. There were 72.8%, 18.6%, and 8.6% of P. falciparum parasites carrying the wild-, mutant-, and mixed-type Pfcrt gene, respectively, and all mutant haplotypes were CVIET (the underline indicates the mutation site). There were 20.0%, 61.4% and 18.6% of P. falciparum parasites carrying the wild-, mutant-, and mixed-type Pfmdr1 gene, respectively, and the mutant haplotypes mainly included YF and NF (the underlines indicate the mutation sites). There were 1.4%, 98.6%, and 0 of P. falciparum parasites carrying the wild-, mutant-, and mixed-type Pfdhfr gene, respectively, and AIRNI was the predominant mutant haplotype (the underline indicates the mutation site). There were 1.4%, 94.3%, and 4.3% of P. falciparum parasites carrying the wild-, mutant-, and mixed-type Pfdhps gene, respectively, and SGKAA was the predominant mutant haplotype (the underline indicates the mutation site). The complete drug-resistant IRNGE genotype consisted of 8.6% of the Pfdhfr and Pfdhps genes, and the K13 gene A578S mutation occurred in 1.4% of the parasite samples. Conclusions There are mutations in the Pfcrt, Pfmdr1, Pfdhfr, Pfdhps, and K13 genes of P. falciparum imported from Equatorial Guinea to Shandong Province, with a low frequency in the Pfcrt gene mutation and a high frequency in the Pfmdr1, Pfdhfr, and Pfdhps gene mutations, and the K13 gene A578S mutation is detected in the parasite samples.

Objective To characterize the epidermal growth factor receptor (EGFR) gene in Schistosoma japonicum (SjEGFR gene) and investigate the role of the EGFR gene in regulating the growth, reproductive system, maturation and fecundity of S. japonicum. Methods Rapid amplification of cDNA ends (RACE) was performed to obtain the full length of the SjEGFR gene, and the SjEGFR gene expression was quantified in different developmental stages of S. japonicum using a quantitative real-time PCR (qPCR) assay. The tissue localization of the SjEGFR gene was detected in 22-day parasite using whole-mount in situ hybridization (WISH). Following RNA interference (RNAi)-induced knockdown of the SjEGFR gene, the worm length, pairing rate and worm burden of S. japonicum were measured, and the worm morphology was observed using optical microscopy and confocal microscopy. Results The SjEGFR gene was identified with a conserved tyrosine-kinase active site, and the SjEGFR gene expression was detected at various developmental stages in male and female parasites. WISH showed that the transcript of the SjEGFR gene was localized on the tegument and in the digestive organs of S. japonicum. RNAi-induced SjEGFR knockdown resulted in marked suppression of the worm growth, smaller size of male testicles that contained more immature spermatocytes, and apparent impairment of ovary and vitelline gland development. In addition, no eggs were found in the uterus of SjEGFR knocked-down female parasites, indicating the interruption of egg production. Conclusions Inhibition of SjEGFR expression may remarkably suppress the growth and maturation of S. japonicum, and interrupt the egg production.