DNA methylation and mRNA expression of COL6A3 in antler mesenchyme of female and male reindeer

Jian‑Cheng Zhai,Ruo‑Bing Han,Sheng‑Nan Wang,Qiang‑Hui Wang,Yan‑Ling Xia,Wei‑Shi Liu,Ya‑Jie Yin,He‑Ping Li 한국유전학회 2019 Genes & Genomics Vol.41 No.9

Backgroud Reindeer is the only deer species that both male and female produce antlers, which provides a particularly interesting case in studying the differences between antlers of the two sexes. Alpha 3(VI) Collagen Gene (COL6A3), forms a microfibrillar network associated with the structural integrity and biomechanical properties, has been found to be one of the differentially expressed genes in antler mesenchyme of female and male reindeer. Objective and Methods The promoter sequence of reindeer COL6A3 gene was obtained using the cloning technology and analyzed by the bioinformatics methods. Bisulfite sequencing PCR (BSP) was used to detect the methylation status of the COL6A3 promoter in reindeer antler mesenchyme. Real-time quantitative PCR was used to detect COL6A3 expression in the antler mesenchyme of female and male reindeer. Results Sequence analysis revealed that the reindeer COL6A3 partial promoter sequence was 983 bp including the possible promoter region at + 105 bp to + 155 bp. Homology and phylogenetic analysis indicated that the COL6A3 promoter of reindeer had the closest genetic distance with Bos taurus, Capra hircus and Ovis aries. BSP results indicated that the methylation level of COL6A3 promoter in the female reindeer antler mesenchyme was significantly higher than in the male. Correlating with increased methylation status, we also found that COL6A3 mRNA expression in female reindeer antler mesenchyme was significantly lower than in the male. Conclusion The higher methylation level of the COL6A3 gene in female reindeer antler mesenchyme coincides with decreased COL6A3 mRNA expression, thereby affecting the transposon silencing mechanism and possibly contributing to apparent differences of antlers in female and male reindeer.

GRADE Methods in traditional medicine

Liu Jian-ping 한국한의학연구원 2022 Integrative Medicine Research Vol.11 No.2

The GRADE (Grading of Recommendation, Assessment, Development and Evaluation) methods was developed to evaluate the quality of evidence and make recommendations, which has been widely adopted in clinical practice guidelines. The GRADE methods address the classification of outcomes, systematic collection, appraisal and synthesis of research evidence based on each outcome, evaluate the overall quality of the evidence, and making recommendations. This essay summarized the GRADE methods and its use in clinical practice guidelines of traditional, complementary, and integrative medicine as well as highlighting some of the challenges.

Analysis of Geographic and Pairwise Distances among Chinese Cashmere Goat Populations

Liu, Jian-Bin,Wang, Fan,Lang, Xia,Zha, Xi,Sun, Xiao-Ping,Yue, Yao-Jing,Feng, Rui-Lin,Yang, Bo-Hui,Guo, Jian Asian Australasian Association of Animal Productio 2013 Animal Bioscience Vol.26 No.3

This study investigated the geographic and pairwise distances of nine Chinese local Cashmere goat populations through the analysis of 20 microsatellite DNA markers. Fluorescence PCR was used to identify the markers, which were selected based on their significance as identified by the Food and Agriculture Organization of the United Nations (FAO) and the International Society for Animal Genetics (ISAG). In total, 206 alleles were detected; the average allele number was 10.30; the polymorphism information content of loci ranged from 0.5213 to 0.7582; the number of effective alleles ranged from 4.0484 to 4.6178; the observed heterozygosity was from 0.5023 to 0.5602 for the practical sample; the expected heterozygosity ranged from 0.5783 to 0.6464; and Allelic richness ranged from 4.7551 to 8.0693. These results indicated that Chinese Cashmere goat populations exhibited rich genetic diversity. Further, the Wright's F-statistics of subpopulation within total (FST) was 0.1184; the genetic differentiation coefficient (GST) was 0.0940; and the average gene flow (Nm) was 2.0415. All pairwise FST values among the populations were highly significant (p<0.01 or p<0.001), suggesting that the populations studied should all be considered to be separate breeds. Finally, the clustering analysis divided the Chinese Cashmere goat populations into at least four clusters, with the Hexi and Yashan goat populations alone in one cluster. These results have provided useful, practical, and important information for the future of Chinese Cashmere goat breeding.

Regulation of matrix metalloproteinase-9 protein expression by 1α , 25-(OH)2D3 during osteoclast differentiation

Jian-Hong Gu,Xi-Shuai Tong,Guohong Chen,Xue-Zhong Liu,Jian-Chun Bian,Yan Yuan,Zong-Ping Liu 대한수의학회 2014 Journal of Veterinary Science Vol.15 No.1

To investigate 1α,25-(OH)2D3 regulation of matrixmetalloproteinase-9 (MMP-9) protein expression duringosteoclast formation and differentiation, receptor activator ofnuclear factor κB ligand (RANKL) and macrophagecolony-stimulating factor (M-CSF) were administered toinduce the differentiation of RAW264.7 cells into osteoclasts. The cells were incubated with different concentrations of1α,25-(OH)2D3 during culturing, and cell proliferation wasmeasured using the methylthiazol tetrazolium method. Osteoclast formation was confirmed using tartrate-resistantacid phosphatase (TRAP) staining and assessing bone lacunarresorption. MMP-9 protein expression levels were measuredwith Western blotting. We showed that 1α,25-(OH)2D3inhibited RAW264.7 cell proliferation induced by RANKLand M-CSF, increased the numbers of TRAP-positiveosteoclasts and their nuclei, enhanced osteoclast boneresorption, and promoted MMP-9 protein expression in aconcentration-dependent manner. These findings indicatethat 1α,25-(OH)2D3 administered at a physiological relevantconcentration promoted osteoclast formation and couldregulate osteoclast bone metabolism by increasing MMP-9protein expression during osteoclast differentiation.

Structural-based Screening of L-phenylglycine Aminotransferase using L-phenylalanine as the Optimal Amino Donor: Recycling of L-phenylalanine to Produce L-phenylglycine

Shuang Ping Liu,Rui Xia Liu,Jian Mao,Liang Zhang,Zhongyang Ding,Zhenghua Gu,Gui-Yang Shi 한국생물공학회 2016 Biotechnology and Bioprocess Engineering Vol.21 No.1

The aproteinogenic amino acid, L-phenylglycine, is an important side chain building block for some drugs. It would be of great commercial and environmental value to biocatalyse L-phenylalanine to L-phenylglycine, and thus replace the organic synthesis method. To produce Lphenylglycine from L-phenylalanine, an L-phenylglycine aminotransferase was screened and characterized. HpgTAO showed high homology to α-aminoadipate aminotransferase. The L-phenylalanine binding site was near the residues S26, R401, N201, and G46 in HpgTAO, and L-phenylalanine formed a hydrogen bond with Asn20, which was similar to the substrate binding mechanism of α-aminoadipate aminotransferase. HpgTAO showed increased activity in alkalescent environment below 40°C. The kinetic analysis showed that L-phenylalanine had the highest affinity to HpgTAO, which ensured the recycle biosynthesis of Lphenylglycine from L-phenylalanine. To date, it was the only aminotransferase using L-phenylalanine as an optimal amino donor. The L-phenylglycine biocatalysis operon was also constructed by co-expressing the hmaS, hmo and hpgT by a single plasmid. The first in vitro conversion of Lphenylalanine to L-phenylglycine was achieved by directly using the L-phenylalanine fermentation broth as the raw material.

Outcomes Based on Risk Assessment of Anastomotic Leakage after Rectal Cancer Surgery

Gong, Jian-Ping,Yang, Liu,Huang, Xin-En,Sun, Bei-Cheng,Zhou, Jian-Nong,Yu, Dong-Sheng,Zhou, Xin,Li, Dong-Zheng,Guan, Xin,Wang, Dong-Feng Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.2

Purpose: Anastomotic leakage (AL) is associated with high morbidity and mortality, high reoperation rates, and increased hospital length of stay. Here we investigated the risk factors for AL after anterior resection for rectal cancer with a double stapling technique. Patients and Methods: Data for 460 patients who underwent primary anterior resection with a double stapling technique for rectal carcinoma at a single institution from 2003 to 2007 were prospectively collected. All patients experienced a total mesorectal excision (TME) operation. Clinical AL was defined as the presence of leakage signs and confirmed by diagnostic work-up according to ICD-9 codes 997.4, 567.22 (abdominopelvic abscess), and 569.81 (fistula of the intestine). Univariate and logistic regression analyses of 20 variables were undertaken to determine risk factors for AL. Survival was analysed using the Cox regression method. Results: AL was noted in 35 (7.6%) of 460 patients with rectal cancer. :Median age of the patients was 65 (50-74) and 161 (35%) were male. The diagnosis of AL was made between the 6th and 12th postoperative day (POD; mean 8th POD). After univariate and multivariate analysis, age (p=0.004), gender (p=0.007), tumor site (p<0.001), preoperative body mass index (EMI) (p<0.001), the reduction of TSGF on 5th POD less than 10U/ml (p=0.044) and the pH value of pelvic dranage less than or equal to 6.978 on 3rd POD (p<0.001) were selected as 6 independent risk factors for AL. It was shown that significant differences in survival of the patients were AL-related (p<0.001), high ASA score related (p=0.036), high-level EMI related (p=0.007) and advanced TNM stage related <p<0.001). Conclusions: AL after anterior resection for rectal carcinoma is related to advanced age, low tumor site, male sex, high preoperative EMI, low pH value of pelvic drainage on POD 3 and a significant reduction of TSGF on POD 5. In addition to their high risk of immediate postoperative morbidity and mortality, AL, worse physical status, severe obesity and advanced TNM stage have similarly negative impact on survival.

Cadmium induces apoptosis in primary rat osteoblasts through caspase and mitogen-activated protein kinase pathways

Hong-Yan Zhao,Wei Liu,Yi Wang,Nannan Dai,Jian-Hong Gu,Yan Yuan,Xue-Zhong Liu,Jian-Chun Bian,Zong-Ping Liu 대한수의학회 2015 Journal of Veterinary Science Vol.16 No.3

Exposure to cadmium (Cd) induces apoptosis in osteoblasts (OBs); however, little information is available regarding the specific mechanismsof Cd-induced primary rat OB apoptosis. In this study, Cd reduced cell viability, damaged cell membranes and induced apoptosis in OBs. We observed decreased mitochondrial transmembrane potentials, ultrastructure collapse, enhanced caspase-3 activity, and increasedconcentrations of cleaved PARP, cleaved caspase-9 and cleaved caspase-3 following Cd treatment. Cd also increased the phosphorylationof p38-mitogen-activated protein kinase (MAPK), extracellular signal-regulated kinases (ERK)1/2 and c-jun N-terminal kinase (JNK) in OBs. Pretreatment with the caspase inhibitor, N-benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone, ERK1/2 inhibitor (U0126), p38 inhibitor(SB203580) and JNK inhibitor (SP600125) abrogated Cd-induced cell apoptosis. Furthermore, Cd-treated OBs exhibited signs of oxidativestress protection, including increased antioxidant enzymes superoxide dismutase and glutathione reductase levels and decreased formationof reactive oxygen species. Taken together, the results of our study clarified that Cd has direct cytotoxic effects on OBs, which are mediatedby caspase- and MAPK pathways in Cd-induced apoptosis of OBs.

Analysis of Volatile Compounds of Jasmine Scented Tea during the Orthodox Manufacturing Process Using HS-SPME/GC-MS

Jian Lu,De-Jun Fu,Dong-Fen Wu,Yu Liu,Yao-Ping Luo 한국차학회 2015 한국차학회지 Vol.- No.S

Volatile components of jasmine-scented tea during the orthodox manufacturing process were extracted using Headspace solid-phase microextraction (HS-SPME) and the major flavor compounds were identified by Gas chromatography-mass spectrometry/Retention indices (GC-MS/RI). The major volatile compounds extracted by SDE could also be obtained by HS-SPME, which appears more suitable for extraction of aromatic compounds, for its advantages of simplicity and convenience. The GC-MS chromatograms of tea samples showed that the aromatic constituents of jasmine tea had been identified in composition after 1st scenting, which contained 6 apparently high peaks representing linalool, benzyl acetate, α-farnesene, (Z)-3-hexenyl benzoate, methyl anthranilate, and indole. According to the change of peak area of major volatile compounds, (Z)-3-hexenyl acetate and (Z)-3-hexenyl butyrate with green grass aroma reduced by 71.13% and 63.10% respectively in jasmine tea product, while 7 compounds with favorable flowery fruity aroma-benzyl acetate, (Z)-3-hexenyl benzoate, linalool, benzyl alcohol, α-farnesene, methyl anthranilate and indole- increased by 17.84%, 28.33 times, 32.49%, 1.70 times, 6.67 times, 11.84 times and 21.48 times respectively, comparing to the base tea. Besides, nerolidol (fruity-flowery) and germacrene D (woody), not detected in the base tea also gradually increased. The quality grade of jasmine tea was potentially correlated to the ratio of peak area of the 9 favorable compounds to that of the 2 grass compounds. After 1st scenting, the peak areas of compounds from jasmine flower increased by 1.81×105-18.24×105, and decreased by 36.14%-69.95% after the 1st drying. The peak areas of some components detected in both the base tea and jasmine flowers, like (Z)-3-hexenyl acetate and methyl salicylate, increased by 9.05%-57.60% of those of the base tea after 1st scenting, and lost more than 100% after the 1st drying, while the increased peak areas of some other components both detected, like α-farnesene and indole, were more than peak areas of the base tea, with 30%- 95% loss after the 1st drying. The content change tendency of volatiles during 2nd scenting and the 2nd drying were consistent with those during 1st scenting and the 1st drying. After raising fragrance, the contents of some low boiling point volatiles and compounds not detected in the base tea increased.

SELECTIVE BREEDING IN SWIMMING CRAB (Portunus trituberculatus)

Jian Li,Ping Liu,Baoquan Gao 한국수산과학회 양식분과 2015 한국수산과학회 양식분과 학술대회 Vol.2015 No.5

Clinicopathologic Characteristics and Prognostic Factors in Patients with Operable HER-2 Overexpressing Breast Cancer

Liu, Ai-Na,Sun, Ping,Liu, Jian-Nan,Ma, Jin-Bo,Qu, Hua-Jun,Zhu, Hua,Yu, Cai-Yan,Zhang, Liang-Ming Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.4

Objective: To study the relationship between clinical pathologic characteristics, treatment modalities and prognostic factors in HER-2 (Human Epidermal growth factor Receptor-2) overexpressed breast carcinoma. Materials and Methods: Major clinico-pathological factors including therapeutic modalities and survival status of 371 breast cancer patients with HER2 over-expression, teated at Yantai Yuhuangding Hospital from March of 2002 to December of 2010 were retrospectively studied, with special attention focused on survival-related factors. Results: The median age of the total 371 patients in this study was 48 years at time of diagnosis, among which, the leading pathological type was infiltrating ductal carcinoma (92.5%); 62.8% presented with a primary tomor larger than 2 cm in diameter at diagnosis, 51.0% had axillary lymph node (ALN) metastases; ER (Estrogen receptor)/PR (Progesterone receptor) double negative occured in 52.8% of cases, and PCNA (proliferation cell nuclear antigen) (+++) was found in 55.1%. HER-2 overexpressed patients were usually in advanced stage when the diagnosis was made (72.8% at stages IIA~IIIC). The prognosis and survival were assessed in 259 patients with complete follow-up data. 5-year DFS (disease-free survival) and OS (overall survival) rate was 68.0% and 78.0% respectively. Univariate analysis revealed that age, tumor size, ALN metastases, LVSI (lymph-vascular space involvement), PCNA status, hormonal therapy, chemotherapy cycles, and HER-2 overexpression, correlated closely with the prognosis. ALN metastases, LVSI, PCNA status and chemotherapy cycles were independent predictors of survival. Conclusions: HER-2 overexpressed breast cancer has special clinical and pathological characteristics, with advanced clinical stages and high rate of ER/PR double negative. Lymph node metastases, LVSI, PCNA and chemotherapy cycles are independent predictors of prognosis.