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Disease Progression from Chronic Hepatitis C to Cirrhosis and Hepatocellular Carcinoma is Associated with Increasing DNA Promoter Methylation

  • Zekri, Abd El-Rahman Nabawy;Nassar, Auhood Abdel-Monem;El-Rouby, Mahmoud Nour El-Din;Shousha, Hend Ibrahim;Barakat, Ahmed Barakat;El-Desouky, Eman Desouky;Zayed, Naglaa Ali;Ahmed, Ola Sayed;Youssef, Amira Salah El-Din;Kaseb, Ahmed Omar;El-Aziz, Ashraf Omar Abd;Bahnassy, Abeer Ahmed
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.11
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    • pp.6721-6726
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    • 2013
  • Background: Changes in DNA methylation patterns are believed to be early events in hepatocarcinogenesis. A better understanding of methylation states and how they correlate with disease progression will aid in finding potential strategies for early detection of HCC. The aim of our study was to analyze the methylation frequency of tumor suppressor genes, P14, P15, and P73, and a mismatch repair gene (O6MGMT) in HCV related chronic liver disease and HCC to identify candidate epigenetic biomarkers for HCC prediction. Materials and Methods: 516 Egyptian patients with HCV-related liver disease were recruited from Kasr Alaini multidisciplinary HCC clinic from April 2010 to January 2012. Subjects were divided into 4 different clinically defined groups - HCC group (n=208), liver cirrhosis group (n=108), chronic hepatitis C group (n=100), and control group (n=100) - to analyze the methylation status of the target genes in patient plasma using EpiTect Methyl qPCR Array technology. Methylation was considered to be hypermethylated if >10% and/or intermediately methylated if >60%. Results: In our series, a significant difference in the hypermethylation status of all studied genes was noted within the different stages of chronic liver disease and ultimately HCC. Hypermethylation of the P14 gene was detected in 100/208 (48.1%), 52/108 (48.1%), 16/100 (16%) and 8/100 (8%) among HCC, liver cirrhosis, chronic hepatitis and control groups, respectively, with a statistically significant difference between the studied groups (p-value 0.008). We also detected P15 hypermethylation in 92/208 (44.2%), 36/108 (33.3%), 20/100 (20%) and 4/100 (4%), respectively (p-value 0.006). In addition, hypermethylation of P73 was detected in 136/208 (65.4%), 72/108 (66.7%), 32/100 (32%) and 4/100 (4%) (p-value <0.001). Also, we detected O6MGMT hypermethylation in 84/208 (40.4%), 60/108 (55.3%), 20/100 (20%) and 4/100 (4%), respectively (p value <0.001. Conclusions: The epigenetic changes observed in this study indicate that HCC tumors exhibit specific DNA methylation signatures with potential clinical applications in diagnosis and prognosis. In addition, methylation frequency could be used to monitor whether a patient with chronic hepatitis C is likely to progress to liver cirrhosis or even HCC. We can conclude that methylation processes are not just early events in hepatocarcinogenesis but accumulate with progression to cancer.

Changes of Leaf Area Index, Physiological Activities and Soil Water in Tricholoma matsutake Producing Pine Forest Ecosystem (송이산 소나무림 생태계에서 엽면적지수와 생리적활동 및 토양수분의 변화)

  • Koo, Chang-Duck;Ka, Kang-Hyun;Park, Won-Chul;Park, Hyun;Ryu, Sung-Ryul;Park, Yong-Woo;Kim, Tae-Heon
    • Journal of Korean Society of Forest Science
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    • v.96 no.4
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    • pp.438-447
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    • 2007
  • The purpose of this study was to understand the ecosystem structure and function and soil water changes in Tricholoma matsutake producing pine stands. The investigated stands were pine forest in Sogrisan National Park in Chungbuk-do of Korea. For the purpose we investigated main vegetation, leaf area index(LAI) as ecosystem structural factors and measured photosynthesis, transpiration, xylem water potential, and soil water changes as ecosystem functional factors. Vertical vegetation structure of the site was composed of Pinus densiflora as a overstory species, Quercus mongolica as midstory, Rhododendron mucronulatum, R. schlippenbachii and Fraxinus sieboldiana as understory ones. In the stands LAI was 3.8 during June to September, 2.6 in October and 2.1 during November to April. Photosyntheses of the trees were 6.0 to $7.0{\mu}mol\;CO_2/m^2/s$ in August, and for P. densiflora about $4.0{\mu}mol\;CO_2/m^2/s$ and for Q. mongolica $2.0{\mu}mol\;CO_2/m^2/s$ in mid October. However, R. mucronulatum stopped fixing $CO_2$ and F. sieboldiana shed off the leaves already in mid October. Transpirations were 2.5 to $3.5mmol\;H_2O/m^2/s$ in late August and about $1.0mmol/H_2O/m^2/s$ in mid October. Plant water potentials were -10 to -22 bars for P. densiflora and -5 to -12 bars for the other woody species. The lowest potentials was in late August and highest in late October. Soil water in the stand was closely related to topography. Soil water contents were 7 to 11% at the ridge, 8 to 15% at the hillside and 11 to 19% at the base. Soil temperatures were 0.2 to $0.4^{\circ}C$ higher in T. matustuake colony than noncolony. Mid September soil temperature decreased to $19^{\circ}C$ at which T. matsutake forms primordia. In T. matsutake colony soil moisture was 0.5 to 2.0% lower due to metabolism for consuming water. We suggest that the complicate relationships between ecosystem structure and function in Tricholoma matsutake producing pine stand need to be further investigated.

Effects of Ginsenosides and Their Metabolites on Voltage-dependent Ca2+ Channel Subtypes

  • Lee, Jun-Ho;Jeong, Sang Min;Kim, Jong-Hoon;Lee, Byung-Hwan;Yoon, In-Soo;Lee, Joon-Hee;Choi, Sun-Hye;Lee, Sang-Mok;Park, Yong-Sun;Lee, Jung-Ha;Kim, Sung Soo;Kim, Hyoung-Chun;Lee, Boo-Yong;Nah, Seung-Yeol
    • Molecules and Cells
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    • v.21 no.1
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    • pp.52-62
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    • 2006
  • In previous reports we demonstrated that ginsenosides, active ingredients of Panax ginseng, affect some subsets of voltage-dependent $Ca^{2+}$ channels in neuronal cells expressed in Xenopus laevis oocytes. However, the major component(s) of ginseng that affect cloned $Ca^{2+}$ channel subtypes such as ${\alpha}_{1C}$(L)-, ${\alpha}_{1B}$(N)-, ${\alpha}_{1A}$(P/Q)-, ${\alpha}_{1E}$(R)- and ${\alpha}_{1G}$(T) have not been identified. Here, we used the two-microelectrode voltage clamp technique to characterize the effects of ginsenosides and ginsenoside metabolites on $Ba^{2+}$ currents ($I_{Ba}$) in Xenopus oocytes expressing five different $Ca^{2+}$ channel subtypes. Exposure to ginseng total saponins (GTS) induced voltage-dependent, dose-dependent and reversible inhibition of the five channel subtypes, with particularly strong inhibition of the ${\alpha}_{1G}$-type. Of the various ginsenosides, $Rb_1$, Rc, Re, Rf, $Rg_1$, $Rg_3$, and $Rh_2$, ginsenoside $Rg_3$ also inhibited all five channel subtypes and ginsenoside $Rh_2$ had most effect on the ${\alpha}_{1C}$- and ${\alpha}_{1E}$-type $Ca^{2+}$ channels. Compound K (CK), a protopanaxadiol ginsenoside metabolite, strongly inhibited only the ${\alpha}_{1G}$-type of $Ca^{2+}$ channel, whereas M4, a protopanaxatriol ginsenoside metabolite, had almost no effect on any of the channels. $Rg_3$, $Rh_2$, and CK shifted the steady-state activation curves but not the inactivation curves in the depolarizing direction in the ${\alpha}_{1B}$- and ${\alpha}_{1A}$-types. These results reveal that $Rg_3$, $Rh_2$ and CK are the major inhibitors of $Ca^{2+}$ channels in Panax ginseng, and that they show some $Ca^{2+}$ channel selectivity.

A study of the plan dosimetic evaluation on the rectal cancer treatment (직장암 치료 시 치료계획에 따른 선량평가 연구)

  • Jeong, Hyun Hak;An, Beom Seok;Kim, Dae Il;Lee, Yang Hoon;Lee, Je hee
    • The Journal of Korean Society for Radiation Therapy
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    • v.28 no.2
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    • pp.171-178
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    • 2016
  • Purpose : In order to minimize the dose of femoral head as an appropriate treatment plan for rectal cancer radiation therapy, we compare and evaluate the usefulness of 3-field 3D conformal radiation therapy(below 3fCRT), which is a universal treatment method, and 5-field 3D conformal radiation therapy(below 5fCRT), and Volumetric Modulated Arc Therapy (VMAT). Materials and Methods : The 10 cases of rectal cancer that treated with 21EX were enrolled. Those cases were planned by Eclipse(Ver. 10.0.42, Varian, USA), PRO3(Progressive Resolution Optimizer 10.0.28) and AAA(Anisotropic Analytic Algorithm Ver. 10.0.28). 3fCRT and 5fCRT plan has $0^{\circ}$, $270^{\circ}$, $90^{\circ}$ and $0^{\circ}$, $95^{\circ}$, $45^{\circ}$, $315^{\circ}$, $265^{\circ}$ gantry angle, respectively. VMAT plan parameters consisted of 15MV coplanar $360^{\circ}$ 1 arac. Treatment prescription was employed delivering 54Gy to recum in 30 fractions. To minimize the dose difference that shows up randomly on optimizing, VMAT plans were optimized and calculated twice, and normalized to the target V100%=95%. The indexes of evaluation are D of Both femoral head and aceta fossa, total MU, H.I.(Homogeneity index) and C.I.(Conformity index) of the PTV. All VMAT plans were verified by gamma test with portal dosimetry using EPID. Results : D of Rt. femoral head was 53.08 Gy, 50.27 Gy, and 30.92 Gy, respectively, in the order of 3fCRT, 5fCRT, and VMAT treatment plan. Likewise, Lt. Femoral head showed average 53.68 Gy, 51.01 Gy and 29.23 Gy in the same order. D of Rt. aceta fossa was 54.86 Gy, 52.40 Gy, 30.37 Gy, respectively, in the order of 3fCRT, 5fCRT, and VMAT treatment plan. Likewise, Lt. Femoral head showed average 53.68 Gy, 51.01 Gy and 29.23 Gy in the same order. The maximum dose of both femoral head and aceta fossa was higher in the order of 3fCRT, 5fCRT, and VMAT treatment plan. C.I. showed the lowest VMAT treatment plan with an average of 1.64, 1.48, and 0.99 in the order of 3fCRT, 5fCRT, and VMAT treatment plan. There was no significant difference on H.I. of the PTV among three plans. Total MU showed that the VMAT treatment plan used 124.4MU and 299MU more than the 3fCRT and 5fCRT treatment plan, respectively. IMRT verification gamma test results for the VMAT plan passed over 90.0% at 2mm/2%. Conclusion : In rectal cancer treatment, the VMAT plan was shown to be advantageous in most of the evaluation indexes compared to the 3D plan, and the dose of the femoral head was greatly reduced. However, because of practical limitations there may be a case where it is difficult to select a VMAT treatment plan. 5fCRT has the advantage of reducing the dose of the femoral head as compared to the existing 3fCRT, without regard to additional problems. Therefore, not only would it extend survival time but the quality of life in general, if hospitals improved radiation therapy efficiency by selecting the treatment plan in accordance with the hospital's situation.

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Neuroprotective and Antioxidant Effects of Novel Benzofuran-2-Carboxamide Derivatives

  • Cho, Jungsook;Park, Chowee;Lee, Youngmun;Kim, Sunyoung;Bose, Shambhunath;Choi, Minho;Kumar, Arepalli Sateesh;Jung, Jae-Kyung;Lee, Heesoon
    • Biomolecules & Therapeutics
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    • v.23 no.3
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    • pp.275-282
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    • 2015
  • In the present study, we synthesized a series of novel 7-methoxy-N-(substituted phenyl)benzofuran-2-carboxamide derivatives in moderate to good yields and evaluated their neuroprotective and antioxidant activities using primary cultured rat cortical neuronal cells and in vitro cell-free bioassays. Based on our primary screening data with eighteen synthesized derivatives, nine compounds (1a, 1c, 1f, 1i, 1j, 1l, 1p, 1q and 1r) exhibiting considerable protection against the NMDA-induced excitotoxic neuronal cell damage at the concentration of $100{\mu}M$ were selected for further evaluation. Among the selected derivatives, compound 1f (with $-CH_3$ substitution at R2 position) exhibited the most potent and efficacious neuroprotective action against the NMDA-induced excitotoxicity. Its neuroprotective effect was almost comparable to that of memantine, a well-known NMDA antagonist, at $30{\mu}M$ concentration. In addition to 1f, compound 1j (with -OH substitution at R3 position) also showed marked anti-excitotoxic effects at both 100 and $300{\mu}M$ concentrations. These findings suggest that $-CH_3$ substitution at R2 position and, to a lesser degree, -OH substitution at R3 position may be important for exhibiting neuroprotective action against excitotoxic damage. Compound 1j was also found to scavenge 1,1-diphenyl-2-picrylhydrazyl radicals and inhibit in vitro lipid peroxidation in rat brain homogenate in moderate and appreciable degrees. Taken together, our structure-activity relationship studies suggest that the compound with $-CH_3$ substitution at R2 and -OH substitution at R3 positions of the benzofuran moiety might serve as the lead exhibiting potent anti-excitotoxic, ROS scavenging, and antioxidant activities. Further synthesis and evaluation will be necessary to confirm this possibility.

Isolation and Characterization of Mouse Testis Specific Serine/Threonine Kinase 5 Possessing Four Alternatively Spliced Variants

  • Wei, Youheng;Fu, Guolong;Hu, Hairong;Lin, Gang;Yang, Jingchun;Guo, Jinhu;Zhu, Qiquan;Yu, Long
    • BMB Reports
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    • v.40 no.5
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    • pp.749-756
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    • 2007
  • Phosphorylation on serine/threonine or tyrosine residues of target proteins is an essential and significant regulatory mechanism in signal transduction during many cellular and life processes, including spermatogenesis, oogenesis and fertilization. In the present work, we reported the isolation and characterization of mouse testis-specific serine/threonine kinase 5 (Tssk5), which contains four alternatively spliced variants including, Tssk5$\alpha$, Tssk5$\beta$, Tssk5$\gamma$ and Tssk5$\delta$. Moreover, the locus of Tssk5 is on chromosome 14qC3 and the four variants had a similar high expression in the testis and the heart; however, had a low expression in other tissues, except for Tssk5$\alpha$ which also had comparably high expression in the spleen. Each variant of Tssk5 expression began in the testis 16 days after birth. Aside from TSSK5$\alpha$, the other isoforms have an insertion of ten amino acid residues (RLTPSLSAAG) in region VIb (HRD domain) (His-Arg-Asp). Moreover, only TSSK5$\alpha$ exhibited kinase activity and consistently, a further Luciferase Reporter Assay demonstrated that TSSK5$\beta$, TSSK5$\gamma$ and TSSK5$\delta$ cannot be stimulated at the CREB/CRE responsive pathway in comparison to TSSK5$\alpha$. These findings suggest that TSSK5$\beta$, TSSK5$\gamma$, TSSK5$\delta$ may be pseudokinases due to the insertion, which may damage the structure responsible for active kinase activity. Pull-down assay experiments indicated that TSSK5$\beta$, TSSK5 $\gamma$ and TSSK5$\delta$ can directly interact with TSSK5$\alpha$. In summary, these four isoforms with similar expression patterns may be involved in spermatogenesis through a coordinative way in testis.

A Study on the Correlation Between Electrical Resistivity and Rock Classification (전기비저항과 암반분류의 상관관계에 대한 고찰)

  • Kwon, Hyoung-Seok;Hwang, Se-Ho;Baek, Hwan-Jo;Kim, Ki-Seog
    • Geophysics and Geophysical Exploration
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    • v.11 no.4
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    • pp.350-360
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    • 2008
  • Electrical resistivity is one of physical property of the earth and measured by electrical resistivity survey, electrical resistivity logging and laboratory test. Recently, electrical resistivity is widely used in determination of rock quality in support pattern design of road and railway tunnel construction sites. To get more reliable rock quality data from electrical resistivity, it needs a lot of test and study on correlation of resistivity and rock quality. Firstly, we did rock property test in laboratory, such as P wave velocity, Young's modulus, uniaxial compressive strength (UCS) and electrical resistivity. We correlate each test results and we found out that electrical resistivity has highly related to P wave velocity, Young's modulus and UCS. Next, we accomplished electrical resistivity survey in field site and carried out electrical resistivity logging at in-situ area. We also performed rock classification, such as RQD, RMR and Q-system and we correlate electrical resistivity to RMR data. We found out that electrical resistivity logging data are highly correlate to RMR. Also we found out that electrical resistivity survey data are lower than electrical resistivity logging data when there are faults or fractures. And it cause electrical resistivity survey data to lowly correlate to RMR.

Impedance Spectroscopy Models for X5R Multilayer Ceramic Capacitors

  • Lee, Jong-Sook;Shin, Eui-Chol;Shin, Dong-Kyu;Kim, Yong;Ahn, Pyung-An;Seo, Hyun-Ho;Jo, Jung-Mo;Kim, Jee-Hoon;Kim, Gye-Rok;Kim, Young-Hun;Park, Ji-Young;Kim, Chang-Hoon;Hong, Jeong-Oh;Hur, Kang-Heon
    • Journal of the Korean Ceramic Society
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    • v.49 no.5
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    • pp.475-483
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    • 2012
  • High capacitance X5R MLCCs based on $BaTiO_3$ ceramic dielectric layers exhibit a single broad, asymmetric arc shape impedance and modulus response over the wide frequency range between 1 MHz to 0.01 Hz. Analysis according to the conventional brick-layer model for polycrystalline conductors employing a series connection of multiple RC parallel circuits leads to parameters associated with large errors and of little physical significance. A new parametric impedance model is shown to satisfactorily describe the experimental spectra, which is a parallel network of one resistor R representing the DC conductivity thermally activated by 1.32 eV, one ideal capacitor C exactly representing bulk capacitance, and a constant phase element (CPE) Q with complex capacitance $A(i{\omega})^{{\alpha}-1}$ with ${\alpha}$ close to 2/3 and A thermally activated by 0.45 eV or ca. 1/3 of activation energy of DC conductivity. The feature strongly indicate the CK1 model by J. R. Macdonald, where the CPE with 2/3 power-law exponent represents the polarization effects originating from mobile charge carriers. The CPE term is suggested to be directly related to the trapping of the electronic charge carriers and indirectly related to the ionic defects responsible for the insulation resistance degradation.

Effects of Alpha-galactosidase Supplementation to Corn-soybean Meal Diets on Nutrient Utilization, Performance, Serum Indices and Organ Weight in Broilers

  • Wang, C.L.;Lu, W.Q.;Li, Defa;Xing, J.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.12
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    • pp.1761-1768
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    • 2005
  • Effects of alpha-galactosidase (GAL) on broiler corn-soybean meal diet was investigated. In experiment 1, sixty cockerels were allocated to five groups, including three enzyme treatments (GAL added at 0, 500, and 1,000 mg/kg diet), a nitrogen-free diet group and a fast group. The true nitrogen-corrected ME (TME$_n$) and true amino acid availability were determined. In experiment 2, 324 day-old chicks were used in a 2${\times}$3 factorial design consisting of two energy contents (high and low) and three GAL levels (0, 250, and 500 mg/kg). Three feeding phases, comprising 0-21 d, 22-35 d and 36-48 d, were involved. GAL addition improved TME$_n$ and the availability of methionine and cystine (p<0.05). The apparent ME (AME) or nitrogen-corrected AME (AME$_n$) and digestibility of dry matter, organic matter, calcium, and phosphorus were improved significantly on d 21, so was crude protein and an interaction of energy and GAL on AME$_n$ (p<0.05) was found on d 35. However, daily intake and daily gain were significantly improved with GAL addition (p<0.05) during 21 d. The small intestine relative weight decreased at 250 mg/kg GAL (p<0.05) on d 35, whereas presented an interaction between GAL and energy on d 21 (p<0.05). Likewise, this treatment increased breast muscle ratio (p<0.05). On d 21, triglycerides level of broilers showed interaction between energy and enzyme levels (p<0.05). Uric acid level in 500 mg/kg GAL declined linearly (p<0.05). On d 35, quadratic effects (p<0.05) were observed in total protein, albumin, globulin and cholesterol content for enzyme supplementation. And the interactive effects of energy and GAL on serum values showed more obviously. The study implies that GAL improved energy and nutrient availability of corn-soybean meal diet in broiler. The GAL supplementation to corn-soybean meal based diet can improve performance of broilers in early stages of growth.

Genetic Diversity of Chinese Indigenous Pig Breeds in Shandong Province Using Microsatellite Markers

  • Wang, J.Y.;Guo, J.F.;Zhang, Q.;Hu, H.M.;Lin, H.C.;Wang, Cheng;Zhang, Yin;Wu, Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.1
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    • pp.28-36
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    • 2011
  • To investigate the genetic diversity of six Chinese indigenous pig breeds in Shandong province (Laiwu Black, Dapulian Black, Licha Black, Yantai Black, Yimeng Black and Wulian Black), explain their genetic relationship and assess their integrity and degree of admixture with three Western commercial breeds (Landrace, Yorkshire and Duroc), 303 individuals from these breeds were genotyped for 26 microsatellite markers. In general, high genetic diversity (observed heterozygosity ranging from 0.5495 to 0.7746) and large breed differentiation ($F_{ST}$ = 0.188) were observed. The indigenous pig breeds in Shandong exhibited consistently higher levels of genetic diversity than the three Western breeds. However, compared with the Western breeds, which have an $F_{ST}$ value of 0.252, the indigenous breeds in Shandong have smaller $F_{ST}$ value of 0.145. The analysis of breed relationship indicated that the six indigenous breeds are classified into two groups. One includes four breeds, Licha, Yantai, Yimeng and Wulian, which have experienced large gene introgression of the Western breeds through progressive crossbreeding as well as gene flow among themselves. The other includes Laiwu and Dapulian, which are less influenced by the Western breeds and other indigenous breeds in Shandong in the recent past. The results show that some measures must be taken to effectively protect these indigenous pig breeds in Shandong.