[Natural changes of high-risk HPV in women with negative for intraepithelial lesion or malignancy: a prospective 1 year study].

Objective: To investigate the natural changes of high-risk HPV (HR-HPV) in women with negative for intraepithelial lesion or malignancy (NILM) for regulating HR-HPV screening. Methods: Four hundred and thirty-three newly-diagnosed women were enrolled from January 1st, 2015 to December 31, 2015 in Beijing Obstetrics and Gynecology Hospital. The ages of these patients were between 22 and 74 years, the average age was (45±21) years old. Two hundred and sixty-three cases were less than 50 years old, 170 cases were more and equal to 50 years old. One hundred and fifty-six cases were HR-HPV positive, 277 cases were HR-HPV negative. Follow up tests were conducted for all 433 patients, who were screened by ThinPrep cytologic test (TCT) combined with HR-HPV and were diagnosed with NILM, for a period of 1 year (at the 3, 6, 9 and 12 months intervals respectively), if the TCT results are abnormal and the HR-HPV test results are positive, will follow up colposcopy directed cervical biopsy. Results: (1) HR-HPV natural changes: of 156 NILM cases with HR-HPV infection, 42 cases (26.9%, 42/156) turned negative within 3 months, 88 cases (56.4%, 88/156) turned negative within 6 months, 99 cases (63.5%, 99/156) turned negative within 9 months, and 100 cases (64.1%, 100/156) turned negative within 12 months. The negative conversion ratio at 3, 6, 9 and 12 months for women at childbearing age (<50 years) were significantly higher than those at non-childbearing age (≥50 years old; all P<0.05). Of 277 NILM cases without HR-HPV infection, 35 cases (12.6%, 35/277) had new HR-HPV positive infections within 3 months, 70 cases (25.3%, 70/277) had new infections within 6 months, 80 cases (28.9%, 80/277) had new infections within 9 months, and 83 cases (30.0%, 83/277) had new infections within 12 months. The new infections rate at 3, 6, 9 and 12 months for women at childbearing age (<50 years old) were slightly higher than those at non-childbearing age (≥50 years old; all P>0.05). (2) The progress of cervical leision: of 156 NILM cases with HR-HPV positive, no case progressed during 12 months follow-up. Of 277 NILM cases with HR-HPV negative, 4 cases progressed to cervical intraepithelial neoplaisa (CIN) with HR-HPV infection and TCT abnormal during 12 months follow-up, including 2 cases pathology diagnosed with CINⅠ, 1 case with CINⅡ, and 1 case with CINⅢ. The progression rate was 1.4%(4/277), which accounts for 4.8% (4/83) of new HR-HPV infections cases in women. Conclusions: The results of cytology combined with HR-HPV screenings suggest every 6 months for simple HR-HPV positive women, colposcopy directed cervical biopsy is recommended to assess cervical lesions if necessary. Cytology combined with HR-HPV screenings suggest every 12 months for simple HR-HPV negative women to early detection of cervical leision.

Nanodroplet microarrays for high-throughput enzyme screening.

We describe here a method for the continuous assessment of enzymatic activity using microarrays. By uniformly coating fluorogenic substrates on slides, we generated surfaces capable of detecting enzymatic activity. The enzymes were deposited on the arrays in segregated droplets using standard microarrayers. Surfaces were developed for assessing the activities of both proteases and phosphatases, hence capitalizing on microarray technology to perform miniaturized high-throughput screens for these, as well as potentially any other, classes of enzyme. This offers an unprecedented ability for performing solution-phase enzymatic assays in nanoliter volumes on microarrays, in contrast to microliter volumes typically required in microplate-based assays, thereby reducing the amounts of reagent(s) required by anywhere from a hundred to a thousand-fold. This new approach thus provides a potentially more cost-effective, label-free enzyme screening technique. A single slide is able to accommodate several thousand assays, facilitating the assessment of both dose and time-dependent inhibition parameters in a single run.