In Vitro Screening for Compounds Derived from Traditional Chinese Medicines with Antiviral Activities Against Porcine Reproductive and Respiratory Syndrome Virus

( Jia Cheng ),( Na Sun ),( Xin Zhao ),( Li Niu ),( Mei Qin Song ),( Yao Gui Sun ),( Jun Bing Jiang ),( Jian Hua Guo2 ),( Yuan Sheng Bai ),( Jun Ping He ),( Hong Quan Li ) 한국미생물 · 생명공학회 2013 Journal of microbiology and biotechnology Vol.23 No.8

Seventeen compounds derived from traditional Chinese medicines (TCMs) were tested for their antiviral activity against porcine reproductive and respiratory syndrome virus (PRRSV) in vitro. Visualization with the cytopathologic effect (CPE) assay and the 3-(4, 5-dimethyithiazol- 2-yl)-2,5-diphenyltetrazolium bromide test were used to determine the 50% cytotoxic concentration (CC50) and 50% effective concentration (EC50) in cultured Marc-145 cells. Among the tested compounds, chlorogenic acid and scutellarin showed potential anti-PRRSV activity. The EC50 values were 270.8 ± 14.6 μg/ml and 28.21 ± 26.0 μg/ml and the selectivity indexes were >5.54 and 35.5, respectively. The time-of-addition and virucidal assay indicated that the anti-PRRSV activity of the two compounds could be due to their inhibiting the early stage of virus replication and/or inactivating the virus directly. The inhibition of the virus attachment was not observed in the adsorption inhibition assay. The inhibition ratios of chlorogenic acid and scutellarin were, respectively, 90.8% and 61.1% at the maximum non-cytotoxic concentrations. The results have provided a basis for further exploration of their antiviral properties and mechanisms in vivo. We believe that the chlorogenic acid and scutellarin have a great potential to be developed as new anti-PRRSV drugs for clinical application.

Cordblood-Based High-Throughput Screening for Deafness Gene of 646 Newborns in Jinan Area of China

Shou-Xia Li,Ding-Li Chen,Su-Bin Zhao,Li-Li Guo,Hai-Qin Feng,Xiao-Fang Zhang,Li-Li Ping,Zhi-Ming Yang,Cai-Xia Sun,Gen-Dong Yao 대한이비인후과학회 2015 Clinical and Experimental Otorhinolaryngology Vol.8 No.3

Objectives. Infants with slight/mild or late-onset hearing impairment might be missed in universal newborn hearing screening (UNHS). We identified the mutation hot spot of common deaf gene in the newborns in Jinan area population by screening the mutation spot with neonate cord blood, in order to make clear whether the neonate cord blood for screening is feasible. Methods. Six hundred and forty-six newborns were subjected to both UNHS and genetic screening for deafness by using neonate cord blood. The newborn genetic screening targeted four deafness-associated genes, which were commonly found in the Chinese population including gap junction beta-2 protein (GJB2), gap junction beta-3 protein (GJB3), solute carrier family 26 member 4 (SLC26A4), and mtDNA 12S rRNA. The most common 20 spot mutations in 4 deaf genes were detected by MassARRAY iPLEX platform and mitochondrial 12S rRNA A1555G and C1494T mutations were sequenced using Sanger sequencing. Results. Among the 646 newborns, 635 cases passed the UNHS and the other 11 cases (1.7%) did not. Of the 11 failures, two cases were found to carry homozygous GJB2 p.R143W pathogenic mutation, one case was found to have heterozygous GJB2 235delC mutation, and another one case carried heterozygous GJB3 p.R180X pathogenic mutation. Six hundred and thirty-five babies passed the newborn hearing screening, in which 25 babies were identified to carry pathogenic mutations, including 12 heterozygotes (1.9%) for GJB2 235delC, eight heterozygotes (1.3%) for SLC26A4 IVS7-2A>G, one heterozygote (0.2%) for p.R409H, two homozygotes (0.3%) for m.1494C>T, and two homozygotes (0.3%) for m.1555A>G. Conclusion. Newborn genetic screening through the umbilical cord blood for common deafness-associated mutations may identify carriers sensitive to aminoglycoside antibiotic, and can effectively prevent or delay hearing loss occurs.

The Catechol-O-Methyltransferase Val158Met Polymorphism Contributes to the Risk of Breast Cancer in the Chinese Population: An Updated Meta- Analysis

Guo-Xing Wan,Yu-Wen Cao,Wen-Qin Li,Yu-Cong Li,Feng Li 한국유방암학회 2014 Journal of breast cancer Vol.17 No.2

Purpose: Catechol-O-methyltransferase (COMT) enzyme plays acentral role in estrogen-induced carcinogenesis. Emerging evidencefrom association studies has revealed that the functionalVal158Met polymorphism (rs4680 G>A) of the Catechol-Omethyltransferasegene (COMT) has been implicated in susceptibilityto breast cancer in the Chinese population, while resultsof individual published studies remain inconclusive and inconsistent. To assess this association in the Chinese population, a meta-analysis was performed. Methods: Eligible studies weresearched on MEDLINE, Embase, Cochrane Library, China NationalKnowledge Infrastructure, and the Chinese BiomedicineDatabase. Odds ratios (ORs) with their corresponding 95% confidenceintervals (CIs) were pooled to assess the association betweenCOMT polymorphisms and the risk of breast cancer usingRevMan 5.2 and Stata 12.0 software. Results: The meta-analysisincluded 14 eligible studies, with a total of 4,626 breast cancercases and 5,637 controls. Overall, the COMT Val158Met polymorphism(rs4680 G>A) was significantly associated with an increasedrisk of breast cancer in several genetic models (A/A vs. G/G: OR, 1.59, 95% CI, 1.12–2.27; A/A vs. G/A+G/G: OR, 1.62,95% CI, 1.14–2.29; A vs. G: OR, 1.15, 95% CI, 1.00–1.32), and asubgroup analysis according to menopausal status showed thatthis association was especially evident among premenopausalChinese women (A/A vs. G/G: OR, 1.87, 95% CI, 0.99–3.54; A/Avs. G/A+G/G: OR, 1.94, 95% CI, 1.03–3.63). Conclusion: The resultsof this meta-analysis indicated that COMT Val158Met variantscontribute to breast cancer susceptibility in the Chinesepopulation, particularly among premenopausal women.

Codelivery of hydrophilic and hydrophobic drugs in a microneedle patch for the treatment of skin pigmentation

Li Qin Zhang,Xiao Peng Zhang,Yu Ying Hao,Bao Li Zhang,Xin Dong Guo 한국공업화학회 2020 Journal of Industrial and Engineering Chemistry Vol.88 No.-

When overexposed to ultraviolet B (UVB) irradiation, skin can cause a series of abnormal reactions, suchas excessive pigmentation and solar elastosis. In order to better treat the skin after overexposed to UVB,we designed a dissolving microneedles (MNs) loading hydrophilic ascorbic acid and hydrophobic vitaminA palmitate to achieve codelivery. Here, we confirmed that MNs had enough mechanical strength toinsert the skin and two drugs were delivered subcutaneously in sequence in vitro. In white guinea pigs,surface observation and histological observation results confirmed that two drugs by MNs transdermaldelivery were better than topical administration in the treatment of UVB-damaged skin. The MNs providea method of loading different drugs into the needles and advance the development of transdermal drugdelivery.

Associations Between TLR9 Polymorphisms and Cancer Risk: Evidence from an Updated Meta-analysis of 25,685 Subjects

Wan, Guo-Xing,Cao, Yu-Wen,Li, Wen-Qin,Li, Yu-Cong,Zhang, Wen-Jie,Li, Feng Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.19

A meta-analysis incorporating 34 case-control studies from 19 articles involving 12,197 cases and 13,488 controls was conducted to assess the effects of three genetic variants of Toll-like receptor 9 (TLR9): rs187084, rs352140, and rs5743836. Studies on associations between TLR9 polymorphisms and cancer risk were systematically searched in electronic databases. The reported odds ratios (OR) and 95% confidence intervals (CI) were pooled to assess the strength of any associations. The results showed that the rs187084 polymorphism was significantly associated with an increased risk of cancer (CC vs TC+TT: OR=1.14, 95% CI=1.02-1.28), specifically cervical cancer (C vs T: OR=1.19, 95% CI=1.05-1.34; TC vs TT: OR=1.32, 95% CI=1.10-1.58; CC vs TT: OR=1.31, 95% CI=1.03-1.68; CC+TC vs TT: OR=1.32, 95% CI=1.11-1.56), and that this association was significantly positive in Caucasians (CC vs. TC+TT: OR=1.18, 95% CI=1.01-1.38). The rs352140 polymorphism had a protective effect on breast cancer (GA vs GG: OR=0.77, 95% CI=0.66-0.89), whereas the rs5743836 polymorphism was likely protective for digestive system cancers (CC+TC vs TT: OR=0.81, 95% CI=0.66-0.98). In conclusion, our results suggest that the rs187084 polymorphism may be associated with an elevated cancer risk, whereas polymorphisms of rs352140 and rs5743836 may play protective roles in the development of breast and digestive system cancers, respectively. From the results of this meta-analysis further large-scale case-control studies are warranted to verify associations between TLR9 polymorphisms and cancer.

Analysis of DNA methylation in tissues and development stages of pearl oyster Pinctada fucata

Yao-guo Li,Yun-yan Guan,Qin Li,Mao-xian He 한국유전학회 2015 Genes & Genomics Vol.37 No.3

In this study, we developed a methylationsensitiveamplification polymorphism technique to investigateDNA methylation profiles in different tissues and inthe early-development stages of the pearl oyster Pinctadafucata (P. fucata). Methylation levels in adductor muscle,digestive gland, axe foot, heart, and gill ranged from 11.71to 14.71 %, and significant differences (P\0.05) betweenmethylation levels in different tissues were observed. TheDNA methylation levels of sperm, egg cells, two-cellembryos, morula embryos, trochophore larvae andD-shaped larvae were 13.51, 11.80, 12.14, 12.60, 14.65 and13.18 %, respectively. Development stages of two-cellembryos, morula embryos, trochophore larvae andD-shaped larvae indicated a higher number of identicalDNA methylation status loci in the egg, compared to that inthe sperm. It is probable that DNA methylation patterns ofthe progeny are mainly influenced by the egg, while thesperm may become increasingly important during theprocess of early embryo development. The observeddifferences in methylation levels in the tissues and thedevelopment stages of P. fucata suggest that DNA methylationmay act as an epigenetic regulator during tissuedifferentiation, individual growth, and development.

Species Classification and Quality Assessment of Chaihu (Radix Bupleuri) Based on High-Performance Liquid Chromatographic Fingerprint and Combined Chemometrics Methods

Xiao-Jie Li,Jie Hu,Zhen-Yu Li,Xue-Mei Qin,Li-Zeng Zhang,Xiao-Qing Guo 대한약학회 2011 Archives of Pharmacal Research Vol.34 No.6

A high-performance liquid chromatographic (HPLC) method was established to analyze 36 Chaihu (Radix Bupleuri) samples collected from three species (Bupleurum chinense DC., B. scorzonerifolium Willd. and B. smithii Wolff.). Addition of trifluoroacetic acid into the mobile phase resulted in fingerprint chromatograms with stable baselines. There were thirty-two characteristic peaks in the standard fingerprint of B. chinense DC. Different recognition pattern methods, including similarity analysis (SA), hierarchical cluster analysis (HCA), principal component analysis (PCA) and partial least squares-discrimination analysis (PLS-DA) were utilized to analyze the 36 samples based on the contents of chemical constituents. Consistent results from SA, HCA and PCA analysis illustrated the rationalisation for why B. smithii Wolff. was not quoted in the Chinese Pharmacopoeia and classified samples were in agreement with their species. PLS-DA loading plots showed the chemical markers which had the most influences on the separation among different species. However, SA, HCA and PCA could not differentiate between wild and cultivated B. chinense DC. as well as between samples from different provinces. HPLC fingerprint in combination with chemometric techniques provided a very flexible and reliable method for homogeneity evaluation and quality assessment of traditional Chinese medicine.

Microarray Analysis of Long Non-coding RNA Expression Profile Associated with 5-Fluorouracil-Based Chemoradiation Resistance in Colorectal Cancer Cells

Xiong, Wei,Jiang, Yong-Xin,Ai, Yi-Qin,Liu, Shan,Wu, Xing-Rao,Cui, Jian-Guo,Qin, Ji-Yong,Liu, Yan,Xia, Yao-Xiong,Ju, Yun-He,He, Wen-Jie,Wang, Yong,Li, Yun-Fen,Hou, Yu,Wang, Li,Li, Wen-Hui Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.8

Background: Preoperative 5-fluorouracil (5-FU)-based chemoradiotherapy is a standard treatment for locally advanced colorectal cancer (CRC). However, CRC cells often develop chemoradiation resistance (CRR). Recent studies have shown that long non-coding RNA (lncRNA) plays critical roles in a myriad of biological processes and human diseases, as well as chemotherapy resistance. Since the roles of lncRNAs in 5-FU-based CRR in human CRC cells remain unknown, they were investigated in this study. Materials and Methods: A 5-FU-based concurrent CRR cell model was established using human CRC cell line HCT116. Microarray expression profiling of lncRNAs and mRNAs was undertaken in parental HCT116 and 5-FU-based CRR cell lines. Results: In total, 2,662 differentially expressed lncRNAs and 2,398 mRNAs were identified in 5-FU-based CRR HCT116 cells when compared with those in parental HCT116. Moreover, 6 lncRNAs and 6 mRNAs found to be differentially expressed were validated by quantitative real time PCR (qRT-PCR). Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis for the differentially expressed mRNAs indicated involvement of many, such as Jak-STAT, PI3K-Akt and NF-kappa B signaling pathways. To better understand the molecular basis of 5-FU-based CRR in CRC cells, correlated expression networks were constructed based on 8 intergenic lncRNAs and their nearby coding genes. Conclusions: Changes in lncRNA expression are involved in 5-FU-based CRR in CRC cells. These findings may provide novel insight for the prognosis and prediction of response to therapy in CRC patients.

Anti-proliferative Effects of Atractylis lancea (Thunb.) DC. via Down-regulation of the c-myc/hTERT/Telomerase Pathway in Hep-G2 Cells

Guo, Wei-Qiang,Li, Liang-Zhi,He, Zhuo-Yang,Zhang, Qi,Liu, Jia,Hu, Cui-Ying,Qin, Fen-Ju,Wang, Tao-Yun Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.11

Atractylis lancea (Thunb.) DC. (AL), an important medicinal herb in Asia, has been shown to have anti-tumor effects on cancer cells, but the involved mechanisms are poorly understood. This study focused on potential effects and molecular mechanisms of AL on the proliferation of the Hep-G2 liver cancer cell line in vitro. Cell viability was assessed by MTT test in Hep-G2 cells incubated with an ethanol extract of AL. Then, the effects of AL on apoptosis and cell cycle progression were determined by flow cytometry. Telomeric repeat amplification protocol (TRAP) assays was performed to investigate telomerase activity. The mRNA and protein expression of human telomerase reverse transcriptase (hTERT) and c-myc were determined by real-time RT-PCR and Western blotting. Our results show that AL effectively inhibits proliferation in Hep-G2 cells in a concentrationand time-dependent manner. When Hep-G2 cells were treated with AL after 48h,the $IC_{50}$ was about 72.1 ${\mu}g/mL$. Apoptosis was induced by AL via arresting the cells in the G1 phase. Furthermore, AL effectively reduced telomerase activity through inhibition of mRNA and protein expression of hTERT and c-myc. Hence, these data demonstrate that AL exerts anti-proliferative effects in Hep-G2 cells via down-regulation of the c-myc/hTERT/telomerase pathway.

Prognostic Evaluation of Categorical Platelet-based Indices Using Clustering Methods Based on the Monte Carlo Comparison for Hepatocellular Carcinoma

Guo, Pi,Shen, Shun-Li,Zhang, Qin,Zeng, Fang-Fang,Zhang, Wang-Jian,Hu, Xiao-Min,Zhang, Ding-Mei,Peng, Bao-Gang,Hao, Yuan-Tao Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.14

Objectives: To evaluate the performance of clustering methods used in the prognostic assessment of categorical clinical data for hepatocellular carcinoma (HCC) patients in China, and establish a predictable prognostic nomogram for clinical decisions. Materials and Methods: A total of 332 newly diagnosed HCC patients treated with hepatic resection during 2006-2009 were enrolled. Patients were regularly followed up at outpatient clinics. Clustering methods including the Average linkage, k-modes, fuzzy k-modes, PAM, CLARA, protocluster, and ROCK were compared by Monte Carlo simulation, and the optimal method was applied to investigate the clustering pattern of the indices including platelet count, platelet/lymphocyte ratio (PLR) and serum aspartate aminotransferase activity/platelet count ratio index (APRI). Then the clustering variable, age group, tumor size, number of tumor and vascular invasion were studied in a multivariable Cox regression model. A prognostic nomogram was constructed for clinical decisions. Results: The ROCK was best in both the overlapping and non-overlapping cases performed to assess the prognostic value of platelet-based indices. Patients with categorical platelet-based indices significantly split across two clusters, and those with high values, had a high risk of HCC recurrence (hazard ratio [HR] 1.42, 95% CI 1.09-1.86; p<0.01). Tumor size, number of tumor and blood vessel invasion were also associated with high risk of HCC recurrence (all p< 0.01). The nomogram well predicted HCC patient survival at 3 and 5 years. Conclusions: A cluster of platelet-based indices combined with other clinical covariates could be used for prognosis evaluation in HCC.