Screening of Deletion Variants within the Goat PRDM6 Gene and Its Effects on Growth Traits.

By genome-wide association studies, the PRDM6 gene has been shown to affect multiple, apparently unrelated inherited traits, including bone density and body mass index. Therefore, it is considered a potentially pleiotropic gene. In this study, we identified a 12 bp deletion variant (NC_030814.1:rs651603667, g: 79985625-79985636delTTGACTGATCCA) within the PRDM6 gene in a large sample (SBWC goats; n = 1044). All goat samples were collected in Shaanxi province in July 2018. The frequency of the wt allele was higher than the frequency of the del allele, and this mutation polymorphism confirmed to be consistent with the Hardy-Weinberg equilibrium (p > 0.05). Further results showed that in a group of goats in the yearling period (18 months old, n = 567), this deletion variant of the PRDM6 gene was associated with heart girth (p = 0.027), cannon circumference (p = 0.008), chest depth (p = 2.10 × 10-5), chest width (p = 0.004), body height (p = 0.032), body length (p = 0.044) and hip-width (p = 0.014). For adult SBWC goats (36 months old, n = 477), the effects of the 12 bp variation on growth-related traits were found to make no difference. These findings show that the 12 bp deletion within the goat PRDM6 gene plays an important role in the early growth and development of goats. Using the 12 bp mutation, breeders can quickly and effectively select excellent individual goats at an early stage.

Impact of multidisciplinary collaborative pharmaceutical care on knowledge, adherence, and efficacy of hormone therapy in climacteric women.

BACKGROUND: The objective of this study was to evaluate the impact of pharmaceutical care on the knowledge, adherence, and efficacy of hormone therapy in climacteric women participated in multidisciplinary collaborative clinic, launched by Peking University First Hospital (Beijing, People's Republic of China) in 2012. METHODS: A total of 296 patients were recruited (intervention group n=150, control group n=146). The patients in the intervention group visited the multidisciplinary collaborative clinic for their initial hormone therapy, receiving individualized pharmaceutical care (PC), whereas the control group visited only the general clinic without PC. The pill count method, knowledge assessment questionnaire (Cronbach's α=0.80), and the modified Kupperman Index were used to assess the knowledge, adherence, and efficacy at months 3, 6, and 12. RESULTS: The intervention group, which received PC, showed significantly higher and better knowledge, adherence, and efficacy than the control group, demonstrating the effectiveness of the PC provided. The knowledge scores of the intervention and control groups at months 3, 6, and 12 were (73.12 vs 59.28), (77.63 vs 66.19), and (80.81 vs 66.64); the data for adherence were (90.97% vs 82.17%), (93.21% vs 87.79%), and (95.81% vs 93.38%); and the values of the modified Kupperman Index were (17.15 vs 24.05), (13.22 vs 22.01), and (12.21 vs 23.15), respectively. CONCLUSION: PC improved the knowledge, adherence, and efficacy of hormone therapy in climacteric women. Therefore, the multidisciplinary collaborative model investigated in our study should be advocated in other health care institutions for the benefit of more patients. Further large-sample and long-term studies should be conducted to evaluate the effects of PC on patient clinical outcomes, including its impact on the safety and efficacy of long-term use of hormone therapy, as well as the economic benefits.

Lentiviral CRISPR/Cas9 vector mediated miR-21 gene editing inhibits the epithelial to mesenchymal transition in ovarian cancer cells.

CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats) mediated genome editing is a powerful approach for loss of function studies. Here we report that lentiviral CRISPR/Cas9 vectors are highly efficient in introducing mutations in the precursor miRNA sequence, thus leading to the loss of miRNA expression and function. We constructed four different lentiviral CRISPR/Cas9 vectors that target different regions of the precursor miR-21 sequence and found that these lentiviral CRISPR/Cas9 miR-21 gRNA vectors induced mutations in the precursor sequences as shown by DNA surveyor mutation assay and Sanger sequencing. Two miR-21 lentiviral CRISPR/Cas9 gRNA vectors were selected to probe miR-21 function in ovarian cancer SKOV3 and OVCAR3 cell lines. Our data demonstrate that disruption of pre-miR-21 sequences leads to reduced cell proliferation, migration and invasion. Moreover, CRISPR/Cas9-mediated miR-21 gene editing sensitizes both SKOV3 and OVCAR3 cells to chemotherapeutic drug treatment. Disruption of miR-21 leads to the inhibition of epithelial to mesenchymal transition (EMT) in both SKOV3 and OVCAR3 cells as evidenced by the upregulation of epithelial cell marker E-cadherin and downregulation of mesenchymal marker genes, vimentin and Snai2. The miR-21 target genes PDCD4 and SPRY2 were upregulated in cells transduced with miR-21gRNAs compared to controls. Our study indicates that lentiviral CRISPR/Cas9-mediated miRNA gene editing is an effective approach to address miRNA function, and disruption of miR-21 inhibits EMT in ovarian cancer cells.

[Establishment of reference ranges for thyroid-related indicators in normal pregnant women].

OBJECTIVE: To establish the gestational age-specific reference ranges for thyroid-related indicators of pregnant women in our hospital, especially for serum thyroid stimulating hormone (TSH) in the first trimester. METHODS: A total of 548 pregnant women in the first trimester were collected in Peking University First Hospital from June 2013 to April 2014. Among them, 254 pregnant women with single fetus who conformed to the National Academy of Clinical Biochemistry criteria, without adverse pregnancy outcomes and obstetric complications, were enrolled in the current study. To establish their own self-sequential longitudinal reference intervals, serum TSH, total thyroxine (TT4) and free thyroxine (FT4) levels were measured using Siemens detection kits during three different trimesters. Urine iodine (UI) was measured using digestion instrument in some of the pregnant women. RESULTS: The gestational age-specific reference ranges for thyroid-related indicators of pregnant women in our hospital were as follows: in the first trimester: TSH 0.23-4.08 mU/L, TT4 92.59-186.25 nmol/L, FT4 13.36-20.81 pmol/L; in the second trimester: TSH 0.78-4.25 mU/L, TT4 103.20-180.95 nmol/L, FT4 11.57-16.62 pmol/L; in the third trimester: TSH 0.65-4.52 mU/L, TT4 78.20-174.70 nmol/L, FT4 10.01-20.57 pmol/L. The median level of TSH during 7 to 12 weeks of gestational age was lower than that of 4 to 6 weeks, but with no significant statistical difference (P=0.063). The medians of UI during three trimesters of pregnancy were 211.60 µg/L, 195.55 µg/L and 198.65 µg/L, respectively, which were all classified as adequate iodine status. CONCLUSIONS: The gestational age-specific reference ranges for thyroid-related indicators are different from the kits' reference ranges, which are also different among the three trimesters. It may be more reasonable to establish different reference ranges for thyroid-related indicators at 4 to 6 weeks and 7 to 12 weeks in the first trimester separately.

[Iris recognize-guided LASIK with Zyoptix System for correction of myopic astigmatism].

OBJECTIVE: To assess the safety, efficacy, predictability, stability, and changes in cylindrical degree and axis after iris recognize-guided LASIK with the Zyoptix 4.01 System for the correction of myopic astigmatism. METHODS: The present study was a prospective, nonrandomized, self-controlled trial. Iris recognize-guided LASIK with the Zyoptix 4.01 System (Bausch & Lomb) was performed in 97 patients (183 eyes) with myopic astigmatism in a 6-month trial. Patients were divided into 3 group according to the pre-operative cylindrical degree: Group 1, -0.50 to -1.00D, 79 eyes; Group 2, -1.25 to -2.00D, 70 eyes; Group 3, 2.00 to 4.00D, 34 eyes. They were also grouped by preoperative astigmatism axis: Group A, with the rule (WTR) astigmatism, 126 eyes; Group B, against the rule (ATR), 34 eyes; Group C, oblique axis astigmatism, 23 eyes. After a Hansatome microkeratome cut, iris recognized wavefront-based excimer ablation (Zyoptix 100) was performed. The degree and axis of astigmatism pre-operative and at various times post-operatively (1 day, 1 month, 3 month and 6 months) were analyzed and compared. RESULTS: At 6 month post-operatively, uncorrected visual acuity (VA) was 1.0 or better in 92.3% of the eyes, and 0.5 or better in all eyes. No eye lost > or = 1 lines of best spectacle-corrected VA (BSCVA) at 6 month post-operatively; 60 eyes gained 1 line of BSCVA, and 22 eyes gained 2 lines. The cylindrical degree of astigmatism decreased from (-1.54 +/- 0.65) D of pre-operation to (-0.26 +/- 0.25) D (6 month, post-operatively), the difference between these two groups was statistically significant. The uncorrected VA (UCVA) and the degree of astigmatism returned to normal in the first week and tended to be stable after 1 month. Six months after the operation, WTR astigmatism decreased from106 eyes (per-operation) to 45 eyes (post-operation). ATR astigmatism decreased from 43 eyes (per-operation) to 31 eyes (post-operation). Oblique astigmatism increased from 34 to 38 eyes. Sixty-nine eyes became non-astigmatism 6 months after the operation. CONCLUSION: Iris recognize-guided LASIK using Zyoptix is an effective and safe procedure for the treatment of myopic astigmatism.