Infection Modes and Subtypes of Human Papilloma Virus in Patients.

Objective To investigate the prevalence of human papilloma virus(HPV)subtypes in patients and to provide an evidence for the prevention and treatment of HPV infection and the development of HPV vaccine. Methods Multiplex PCR was used to detect HPV DNA in 6917 patients in Peking Union Medical College Hospital from January 1,2013 to June 30,2015.Totally 5586 patients entered the final analysis after the repeat samples were deleted.The total positive rate of HPV subtypes(including high-risk subtypes including HPV-16,18,31,33,35,39,45,51,52,56,58,59,and 68 and low-risk subtypes including HPV-6 and 11)and the infection status of different age were analyzed. Results The total positive rate of HPV was 36.29%(2027/5586).The positive rate of high-risk subtype was 24.92%(1392/5586)and low-risk subtype was 1.66%(93/5586).The positive rate of multiple was 9.70%(542/5586)and multiple high-risk subtype was 7.75%(433/5586).The positive rate of high-risk subtype and multiple were 25.52%(1366/5353)and 11.16%(26/233)in female and 9.99%(535/5353)and 3.00%(7/233)in male,there were significantly difference(χ2=24.61,χ2=12.45,all P<0.001).The positive rate of low-risk subtypes(3.86%,9/233)in males was significantly higher than that in females(1.57%,84/5353)(χ2=5.84,P=0.007).The high-risk HPV subtype infection mainly was seen in patients aged 31-50 years and the low-risk HPV subtype infection mainly in patients aged 21-40 years.The age of multiple HPV infections from 31-40 years.The lowest turn negative rates of subtype were HPV52 and HPV58.The top three HPV subtypes with the highest positive rates were HPV52,HPV16,and HPV58.Conclusions The positive rates of HPV type are different between male and female patients.The males are mainly infected with low-risk subtypes,whereas the females with high-risk subtypes and the multiple HPV subtypes.The top three high-risk subtypes are HPV-52,16,and 58.HPV subtypes with the lowest secondary negative rates are HPV-52 and 58.HPV infection is mainly seen in young individuals.

[Study on the feasibility of human papilloma virus subtypes detection by automatic nucleic acid extraction workstation].

OBJECTIVE: To verify the feasibility of human papilloma virus(HPV) subtypes detection by AUTRAX automatic nucleic acid extraction workstation. METHODS: A total of 183 HPV test samples (2 562 types) were collected during August 2014 in Peking Union Medical College Hospital. Nucleic acid determination kit of high-risk HPV types (16, 18, 35, 39, 58, 31, 33, 68, 56, 45, 59, 51, 52) and 6 , 11 type (real-time PCR) were applied for detection. Each sample was divided into two parts. One part was treated with manual extraction, which entailed manually preparing PCR reaction system and added the sample to the PCR plate. Another was treated with the AUTRAX automatic nucleic acid extraction workstation, which automatically prepared the reaction system and added to the PCR board. These two parts proceeded to the real-time PCR detection. The result of manual extraction was set as the golden standard and the Kappa consistency analysis was conducted. Meanwhile, precision and pollution prevention of the AUTRAX were verified. RESULTS: The sensitivity of AUTRAX automatic nucleic acids extraction workstation was 97.12% (101/104). The specificity was 99.51% (2 446/2 458). The accuracy was 99.42% (2 547/2 562). The result of Kappa consistency analysis showed that the two parts were the same (Kappa=0.926). Coefficient of variation (CV) of each HPV types was less than 5%.No pollution phenomenon was found. CONCLUSION: AUTRAX automatic nucleic acids extraction workstation can be used for the HPV subtypes detection in clinical settings.

Analysis of gender-specific associations between aldehyde dehydrogenase 2 (ALDH2) rs671 genetic polymorphisms and serum uric acid levels in Han Chinese.

BACKGROUND: Serum uric acid (SUA) is influenced by lifestyle and genetics, and unbalanced SUA levels are linked to various common disorders. While the aldehyde dehydrogenase 2 (ALDH2) rs671 polymorphism appears to be associated with SUA levels, the evidence remains inconclusive. The aim of this study was to examine the distribution of the ALDH2 rs671 polymorphism among Han Chinese in Beijing and determine the association between this polymorphism and SUA. METHODS: A total of 6,461 randomized healthy individuals were included in the study. Biochemical indicators were tested and ALDH2 rs671 polymorphism testing was conducted for subjects enrolled in the study. The distribution of the ALDH2 rs671 polymorphism and the relationship between genotype and the levels of serum lipids and uric acid (UA) were analyzed. RESULTS: The ALDH2 rs671 genotype frequencies were 68.1% (G/G), 29.3% (G/A), and 2.6% (A/A). There was no significant difference in allele distribution between males and females. In males, different ALDH2 genotypes exhibited significant differences in several biochemical analytes, including body mass index (BMI), blood glucose (Glu), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), UA, glutamyl transpeptidase (GGT), and creatinine (Cr) (P<0.05). No such differences were found in females. SUA levels in G/A and A/A-carrying males were significantly lower than those of G/G-carrying males. The effect of the ALDH2 polymorphism on UA was still significant after further adjustment for factors including BMI, Glu, TC, HDL-C, Cr, and GGT. CONCLUSIONS: The ALDH2 polymorphism is related to SUA in Beijing males, and A allele-carrying males have lower SUA levels.

HLA-B*58:01 and rs9263726 have a linkage, but not absolute linkage disequilibrium in Han Chinese population.

HLA-B*58:01 has been demonstrated to be associated with allopurinol-induced severe cutaneous adverse reactions. Since HLA-B*58:01 is too complicated to be identified, it is necessary to select an appropriate surrogate biomarker. In Japan, the rs9263726 allele was considered as a surrogate biomarker for HLA-B*58:01, but this was not the case with the Australian cohort. Due to the conflict results, in this study, we aim to demonstrate whether the rs9263726 allele is a surrogate biomarker for HLA-B*58:01 in Han Chinese population. A total of 353 samples (200 cases from the south and 153 cases from the north) were selected to detect HLA-B*58:01 and rs9263726 allele. The HLA-B*58:01 was identified by sequencing-based method, and the rs9263726 allele was identified by Taqman SNP Genotyping Assays. The results showed that the two alleles had a linkage, but not absolute linkage disequilibrium in Han Chinese population.

Improved sensitivity and specificity for prostate cancer diagnosis based on the urine PCA3/PSA ratio acquired by sequence­specific RNA capture.

Prostate cancer antigen 3 (PCA3) is a non-coding RNA fragment that is overexpressed in prostate cancer cells. However, the clinical applications of PCA3 are highly limited due to the instability of RNA and the lack of reliable and efficient RNA extraction and purification methods. Thus, in the present study, we compared three different methods of RNA extraction to further confirm the higher yield of commercial magnetic beads with poly-T functionalization and a capturer strand. The current protocols for RNA extraction of i) the phenol-chloroform method, ii) the affinity column method and iii) magnetic beads with poly-T functionalization and a capturer strand were applied separately for RNA extraction in urine samples. Reverse transcription­quantitative polymerase chain reaction was performed to evaluate the yield of the three methods of RNA extraction. Furthermore, 52 urine samples after prostate massage from patients suspected of a diagnosis of prostate cancer were collected. The Mag-Cap method and RT-PCR were applied to obtain the PCA3 score. The clinical value of the PCA3 score was investigated by comparison with the pathology of the prostate biopsy. The yield of the Mag-Cap method was higher than that of the phenol­chloroform method and commercial kits. Thirty­four patients were pathologically diagnosed with prostate cancer and 18 with benign prostatic hyperplasia (BPH). It was confirmed that the median PCA3 score was higher among the prostate cancer patients than those with benign disease (53.5 vs. 17, p=0.000). A sensitivity of 82.4% and a specificity of 77.8% were obtained when the cut-off value for the PCA3 score was 28.5. The Mag-Cap method was found to be more efficient for RNA extraction. The urinary PCA3 score is a promising method for prostate cancer screening, detection and diagnosis, and has the potential to reduce unnecessary prostate biopsies.

[Effect of negative pressure suction on rabbit optic nerve and retina].

OBJECTIVE: To study the effect of acute intraocular pressure (IOP) elevation on the optic nerve and retina in laser in situ keratomileusis (LASIK). METHODS: Acute IOP was increased to 65 mm Hg for different period of time (30 s, 1 min and 3 min) by using scleral suction on rabbit eyes. The tissues of retina and optic nerve of immediately extirpated group (instant group), of 2 weeks later extirpated group (recovery group) both after the negative pressure suction and of normal rabbit eyes were examined with electron and light microscope. RESULTS: After the negative pressure suction for 30 s, 1 min, and 3 min respectively, the optic nerve and retinal cells changed lightly; some part of optic nerve and retinal cells changed; optic nerve fibers and retinal cells changed sharply. CONCLUSION: Ultrastructural changes of retina and optic nerve may be induced by acute IOP elevation in LASIK. The longer the time, the more apparent the changes.