Objective To discuss the correlation of the p53 overexpression and human papilloma virus (HPV) infection with the occurrence, development of the vulvar squamous cell carcinoma in elderly women. Methods Samples from 68 patients with vulvar squamous cell carcinoma and samples of formalin-fixed paraffin-embedded blocks from 38 patients with vulvar dystrophy were determined by immunohistochemistry. HPV infection was examined with monoclonal or polyclonal antibodies. Fifteen normal vulvar skin samples were used as controls. Results Positive expression rate of p53 and HPV in vulvar squamous cell carcinoma samples were 54.4% and 45.6% respectively, in the dystrophy samples were 18.4% and 55.3% respectively. All showed significant difference between the two groups. None was positive in the controls. Conclusions There are high expression of p53 and HPV in the elderly women with vulvar squamous cell carcinoma. p53 mutation and high risk HPV infection play an important role in the development of vulvar squamous cell carcinoma.

Objective To establish a detection method integrating DPRA ( direct peptide reactivity assay) with h?CLAT ( human cell line activation test) to screen the skin sensitization potency of chemicals and plant extracts. Methods 12 chemicals and 7 plant extracts were chosen as the test substances. Firstly, the test substances were incubated together with two different peptides ( cysteine and lysine) respectively for reaction for 24 h. The peptide consumptions were analyzed by HPLC. Simultaneously, THP?1 cells were cultured in vitro and then exposed to different concentrations of test sub?stances for 24 h to examine the cell viability, cell surface markers CD54 and CD86 were assessed by flow cytometry. The predicting results were compared further between DPRA and h?CLAT. Results 12 chemicals were distinguished correctly by DPRA classified as 2 non?sensitizers and 10 sensitizers. The results of DPRA were in accordance with h?CLAT. Predic?ting the sensitization potency of plant extracts by DPRA showed that 6 plant extracts were determined as suspected sensiti?zers except for green tea extract. But using the method of h?CLAT, 4 plant extracts were examined as suspected sensitizers except for green tea extract, herba portulacae extract and ginseng fruit extract. The coherence of DPRA and h?CLAT was 0?57. Conclusion This detection method integrating DPRA with h?CLAT can predict single compound accurately. As for complex compound, it can achieve preliminary prediction and need other integrating methods to make a further identifica?tion.

Objective To evaluate advantages of the Herbert screw in treating displaced radial head fractures. Methods The Herbert screw was used to treat 25 segmental fractures of the radial head from since 1991 and the results were compared with those of other treatment methods mentioned in the literature. Results A follow up averaging 6 years and 8 months showed that postoperative function was all excellent or good and that most cases recovered to normal absolutely, without complications. Conclusions The Herbert screw provides such rigid internal fixation for displaced radial head fractures that, after operation, a plaster cast is rarely required and most patients are able to return to work within a few weeks. This method of treatment appears to offer significant advantages over conventional techniques.

Objective To explore the classification of intracranial arachroid cysts(IAC) in CT cisternography(CTC) and its clinicalapplication.Methods 22 cases of IAC diagnosed by plain CT underwent CTC exminaton. IACs were classified into noncommnicatingintracranial arachnoid cyst (NCIAC) and commnicating intracranial arachnoid cyst (CIAC) by wheather or not filled with contrast media in cysts on CTC. NCIAC cases were selected and treated with neuroendoscopic fenestration.Results 15 cases of NCIAC were found by CTC examination. All the NCIAC patients had definite neurologic findings. Postoperatively, all the patients were improved or cured. Follow-upplain CT scan of 9 NCIAC cases showed the cysts were decreased markedly in size, most of the space around the cysts were replaced bynormal cerebral tissue.Conclusion (1)CTC is simple ,safe and specific for making a final diagnosis of IAC. IACs can be classified into CIAC and NCIAC by CTC findings.(2)Neurosurgical indication for IAC is NCIAC patients with symptoms.

Objective To investigate the fetal haemolytic status when IgG antibody's efficiency of pregnant women was ≥1∶512 in blood group incompatibility. Methods We retrospectively analyzed the data of 6 pregnant women whose IgG antibody's efficiency were more than 1∶512 in blood group incompatibility. The levels of bilirubin, IgG antibody's efficiency and blood type in amniotic fluid from amniocentesis were examined. Umbilical blood bilirubin, blood type analysis and Coombs test of newborns were performed. Results Serum IgG antibody's efficiency of pregnant women did not correlate with the severity of haemolytic disease of newborn. Conclusions Fetal haemolytic disease can not be diagnosed only by serum IgG antibody's efficiency of pregnant women.

Objective:To clarify whether the signal peptide of human nerve growth factor can mediate secretory expression of beta-endorphin and whether there is difference between the efficiency of signal peptides from human and mouse nerve growth factor.Methods: Two kinds of eukaryotic vectors containing human or mouse signal sequence-mediated secretory expression of beta-endorphin were constructed.The culture supernatant and cells were collected 48 h after NIH3T3 cells were transfected by the two kinds of vectors,and the cover slips with single-layer cells was prepared.The concentration of beta-endorphin in the culture was determined by radio-immunoassay.The total RNA was extracted from cells and mRNA from fusion genes was assayed by RT-PCR.Cells on cover slips were subjected to immunofluorescence staining.Results: RT-PCR showed that the fusion genes were expressed in NIH3T3 cells;the expression of beta-endorphin was mainly in the cytoplasm of NIH3T3 cells.The concentrations of beta-endorphin in the supernatants 48 h after transfection with pcDNA3.1-hEP and pcDNA3.1-mEP were(280.33?24.16) pg/ml and(191.04?7.96) pg/ml(P

Objective To compare the effects of three kinds of Oncomelania hupensis RNA extraction methods,namely a modified SDS method,TRIzol reagent method,and CTAB method,so as to obtain an economical and efficient method for RNA extraction from O. hupensis. Methods The modified SDS method,TRIzol reagent method and CTAB method were applied to ex-tract the RNA from O. hupensis. A nucleic acid protein analyzer was used to measure the concentration and purity of RNA. The yields were calculated by the concentration of the products. The purity was indicated by A260/A280 and A260/A230. The quality of RNA was inspected by 1% agarose gel electrophoresis. The β-acting gene was selected as the target gene for RT-PCR analysis. Re-sults The RNA yields obtained by using the three kinds of extraction methods were significantly different(F = 16895.85,P <0.01)according to the analysis of variance. The LSD test showed that the yields obtained by using the modified SDS method were the highest,and those obtained by the CTAB method were the lowest. The purity of RNA extracted by the CTAB method was su-perior to that by the other two methods,and the A260/A280 and A260/A230 ratios of the CTAB method were in the range from 1.8-2.0 and 2.0-2.2. The A260/A230 ratios of the other two methods were both lower than 2.0. The RNA extracted by the modified SDS meth-od had the better integrity. The electrophoresis results showed that the 28S rRNA band,18S rRNA band and 5S rRNA band were clear,and there was no obvious smear between each band. The RNA obtained by the TRIzol reagent method had no 28S rRNA band,and that obtained by the CTAB method had no 28S rRNA and 5S rRNA bands. The β-acting gene of the RNA ex-tracted by all the three methods could be amplified by RT-PCR. The costs and time-consuming of the modified SDS method were less than those of the other two methods. Conclusion The modified SDS method is an economic and efficient method,and it is suitable for extracting the RNA of O. hupensis,especially for large sample preparation.

OBJECTIVE: Chengde is a city with many corneal blindness patients In north China. Lacking of corneas donation is the main reason which prevents corneal transplantation. Survey was made by questionnaire in Chengde to evaluate the current situation and the influential factors of corneal donation.METHODS: Survey was made in 3 200 Chengde residents aged 18 years or older, includes outpatients and inpatients of ophthalmology, some undergraduate students and people met accidentally in park, supermarket, station and centre for elders.48.6% are male and 51.4% are female. Self-made questionnaire includes general state, questions about cornea donation and factors influencing cornea donation.RESULTS: Among 3 200 questionnaires, 2 971 were valid. The effective rate was 92.84%. Over 50% people support donating cornea. More than 40% people intend to donate their corneas and support their relatives to donate. Among the factors for not intending to donate cornea, lacking knowledge of cornea donation was the main reason accounting for 42.81%, and worrying about the misusage of donating cornea without corresponding law became the second factor, which accounting for 21.07%. It has no influence on the consciousness of cornea donation by the difference of sex and location between city and countryside.Whereasfession and level of education indeed influence the consciousness of cornea donation, which of the people from 18 to 40 years old was greater than those of over 40, medical workers was greater than those from other fields, the people graduating from secondary specialized school or higher was greater than those graduating under secondary specialized school.CONCLUSION: People in Chengde have a positive attitude towards cornea donation. It is very necessary to enhance the education of cornea donation, establish an easy and smooth way for donation may promote cornea donating. Consummate legislation is also needed for cornea donation.

Objective To screen the mimic epitopes of Rh(D)blood group antigens and identify their immunity from phage display peptide library.Methods A twelve mer phage peptide library was biopanned with anti-Rh(D)monoclonal antibody immobilized on plastic surface.After three round panning,thirty-five clones were randomly selected and positive clones were identified by ELISA and cross-reaction,followed by antibody competition inhibition assay and DNA sequencing to obtain the mimic epitopes of Rh (D)blood type antigens.The target phage clones were characterized and the antigenicity was analyzed by Western blot.Results After the third round screening,phages were enriched,and eleven positive clones were obtained.According to sequencing and competition inhibition analysis,the same"-WP-Q-"structure existed in seven of the eleven clones,and they had more than 40%inhibition ratio.The other clones had no same characteristics with low inhibition ratio possibly due to non-specific binding.Western blot analysis indicated that these phage clones could be specifically recognized by the anti-Rh(D)serum and they shared the same antigenicity of Rh(D)protein.Conclusions Rh(D)mimotope of"-WP-Q-"structure is successfully obtained by phage peptide library screening with anti-Rh(D)monoelonal antibody.The results lay the foundation for further exploration of pathogenesis and vaccine development of Rh(D)hemolytic diseases of newborn.

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