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PKHD1 Gene Silencing May Cause Cell Abnormal Proliferation through Modulation of Intracellular Calcium in Autosomal Recessive Polycystic Kidney Disease

  • Yang, Ji-Yun;Zhang, Sizhong;Zhou, Qin;Guo, Hong;Zhang, Ke;Zheng, Rong;Xiao, Cuiying
    • BMB Reports
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    • v.40 no.4
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    • pp.467-474
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    • 2007
  • Autosomal recessive polycystic kidney disease (ARPKD) is one of the important genetic disorders in pediatric practice. Mutation of the polycystic kidney and hepatic disease gene 1 (PKHD1) was identified as the cause of ARPKD. The gene encodes a 67-exon transcript for a large protein of 4074 amino acids termed fibrocystin, but its function remains unknown. The neoplastic-like in cystic epithelial proliferation and the epidermal growth factor/epidermal growth factor receptor (EGF/EGFR) axis overactivity are known as the most important characteristics of ARPKD. Since the misregulation of $Ca^{2+}$ signaling may lead to aberrant structure and function of the collecting ducts in kidney of rat with ARPKD, present study aimed to investigate the further mechanisms of abnormal proliferation of cystic cells by inhibition of PKHD1 expression. For this, a stable PKHD1-silenced HEK-293T cell line was established. Then cell proliferation rates, intracellular $Ca^{2+}$ concentration and extracellular signal-regulated kinase 1/2 (ERK1/2) activity were assessed after treatment with EGF, a calcium channel blocker and agonist, verapamil and Bay K8644. It was found that PKHD1-silenced HEK-293T cell lines were hyperproliferative to EGF stimulation. Also PKHD1-silencing lowered the intracellular $Ca^{2+}$ and caused EGF-induced ERK1/2 overactivation in the cells. An increase of intracellular $Ca^{2+}$ in PKHD1-silenced cells repressed the EGF-dependent ERK1/2 activation and the hyperproliferative response to EGF stimulation. Thus, inhibition of PKHD1 can cause EGF-induced excessive proliferation through decreasing intracellular $Ca^{2+}$ resulting in EGF-induced ERK1/2 activation. Our results suggest that the loss of fibrocystin may lead to abnormal proliferation in kidney epithelial cells and cyst formation in ARPKD by modulation of intracellular $Ca^{2+}$.

Survival of Food-borne and Pathogenic Microorganisms in Hot Spring Water (온천수에서 식중독 및 병원성 미생물의 생존 양상)

  • Zheng Jian-Bin;Ahn Yong-Sun;Jeong Do-Yeong;Kim Yong-Suk;Shin Dong-Hwa
    • Journal of Food Hygiene and Safety
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    • v.21 no.1
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    • pp.36-40
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    • 2006
  • This study was conducted to investigate the effects of hot spring water against the survival of food-borne and pathogenic microorganisms. Staphylococcus aureus, Bacillus cereus, and Escherichia coli, which are food-borne microorganisms, Candida albicans and Trichophyton mentagrophytes, which are skin disease pathogens, and Helicobacter pylori, gastritis inducing microorganism, were tested. The content of fluoride in tested hot spring water is 14.1 mg/L, which is higher than the standard of safe for drinking water 1.5 mg/L, but the results on 48 other items were up to the standard. Hot spring water didn't show the bactericidal effects against food-borne microorganisms, C. albicans, and H. pylori tested. However, the viable cell populations of B. cereus and T. mentagrophytes were decreased, which were depends on the temperature of hot spring water. From these results, we confirmed that hot spring water didn't show the bactericidal effects against food-borne microorganisms, skin disease pathogens, and gastritis inducing microorganism, but the growth of some microorganisms were inhibited by high temperature ($41^{\circ}C$).

Angiostrongylus cantonensis: Scanning Electron Microscopic Observations on The Cuticle of Moulting Larvae

  • Zeng, Xin;Wei, Jie;Wang, Juan;Wu, Feng;Fung, Feng;Wu, Xiaoying;Sun, Xi;Zheng, Huanqing;Lv, Zhiyue;Wu, Zhongdao
    • Parasites, Hosts and Diseases
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    • v.51 no.6
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    • pp.633-636
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    • 2013
  • Angiostrongylus cantonensis is a parasitic nematode that needs to develop in different hosts in different larval stages. Freshwater snails, such as Pomacea canaliculata, are the intermediate host, and rats are the definitive host. Periodic shedding of the cuticle (moulting) is an important biological process for the survival and development of the parasite in the intermediate and definitive hosts. However, there are few studies on the cuticle alterations between different stages of this parasite. In this study, we observed the ultrastructural appearance and changes of the cuticle of the 2nd/3rd stage larvae (L2/L3) and the 3rd/4th stage larvae (L3/L4) using a scanning electron microscope. We also first divided L2/L3 into late L2 and early L3. The late L2 lacked alae, but possessed a pull-chain-like fissure. Irregular alignment of spherical particles on the cuticle were noted compared to the L3. Alae appeared in the early L3. The old cuticle turned into a thin filmlike structure which adhered to the new cuticle, and spherical particles were seen regularly arranged on the surface of this structure. Regular rectangular cavities were found on the surface of L3/L4. The caudal structure of L3/L4 was much larger than that of L3, but caudal inflation, such as seen in L4, was not observed. These results are the first to reveal the ultrastructural changes of the cuticle of A. cantonensis before and after moulting of L2/L3 and L3/L4.

Seroprevalence and Risk Factors of Fascioliasis in Yaks, Bos grunniens, from Three Counties of Gansu Province, China

  • Zhang, Xiao-Xuan;Feng, Sheng-Yong;Ma, Jian-Gang;Zheng, Wen-Bin;Yin, Ming-Yang;Qin, Si-Yuan;Zhou, Dong-Hui;Zhao, Quan;Zhu, Xing-Quan
    • Parasites, Hosts and Diseases
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    • v.55 no.1
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    • pp.89-93
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    • 2017
  • The aim of this study was to determine the seroprevalence and risk factors of fascioliasis in yaks, Bos grunniens, from 3 counties of Gansu Province in China. A total of 1,584 serum samples, including 974 samples from white yaks from Tianzhu, 464 from black yaks from Maqu, and 146 from black yaks from Luqu County, were collected and analyzed using ELISA to detect IgG antibodies against Fasciola hepatica. The overall F. hepatica seroprevalence was 28.7% (454/1,584), with 29.2% in white yaks (284/974) and 27.9% in black yaks (170/610). The seroprevalence of F. hepatica in yaks from Tianzhu, Luqu, and Maqu was 29.2%, 22.6%, and 29.5%, respectively. Female yaks (30.9%) had higher F. hepatica seroprevalence than male yaks (23.4%). Also, F. hepatica seroprevalence varied by different age group from 24.1% to 33.8%. Further, the seroprevalence ranged from 21.8% to 39.1% over different seasons. Interestingly, the season and age of yaks were associated with F. hepatica infection in yaks in the investigated areas. These findings provided a basis for further studies on this disease in yaks from 3 counties of Gansu Province in northwestern China, which may ultimately support the development of effective control strategies of fascioliasis in these areas.

Catabolic Enzyme Activities and Physiological Functionalities of Lactic Acid Bacteria Isolated from Korean Traditional Meju (재래식 메주에서 분리한 유산균들의 각종 효소활성 및 기능성)

  • Jeong, Ji-Kang;Zheng, Yanfei;Choi, Hye-Sun;Han, Gwi-Jung;Park, Kun-Young
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.12
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    • pp.1854-1859
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    • 2010
  • Three kinds of Korean traditional Meju were selected and the counts of microorganisms in these Meju were determined. The counts of total aerobic bacteria, lactic acid bacteria and yeasts and molds were 107~108, 106~108 and 107~108 cfu/g, respectively in three Meju and lactic acid bacteria were important microorganisms in the fermentation of Meju. Therefore, we isolated three kinds of dominant lactic acid bacteria from these Meju. They were identified as Leuconostoc mesenteroides (98%, Lm-SMm), Lactobacillus plantarum (99%, Lp-SMm) and Lactococcus lactis (98%, Ll-GAm). Then, enzyme activities and physiological functionalities of three lactic acid bacteria were investigated. Protease, lipase and $\alpha$-amylase activities were detected in three lactic acid bacteria, Ll-GAm showed relatively higher activities than other two lactic acid bacteria. Lm-SMm, Lp-SMm and Ll-GAm showed 45, 48 and 60% of antioxidative activity to 1,1-diphenyl-2-picryhydrazyl (DPPH), and exhibited 45, 67 and 70% of inhibitory effects in HT-29 human colon cancer cells, respectively. These results indicate that three lactic acid bacteria isolated from traditional Meju, especially Ll-GAm are applicable to Meju preparation for soybean paste industry.

Effects of Vanadium Water on Cancer-related Fatigue of Non Advanced Cancer Patients: Randomized Double Blinded Controlled Trial (비진행성 암환자의 암성 피로에 대한 바나듐수의 효능: 무작위배정 이중맹검 대조군 시험)

  • Zheng, Hong-Mei;Kim, Dong-Hee;Lee, Suk-Hoon;Park, Yoo-Kyoung;Yoo, Hwa-Seung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.12
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    • pp.1800-1806
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    • 2010
  • This study was conducted to evaluate the efficacy and safety of vanadium water (VW) on cancer-related fatigue of non-advanced cancer patients. For the experimental group administration, participants consumed three 500 mL/bottle bottles of Jeju Island's VW, which contained 15~20 ppb of vanadium, three times a day (morning, afternoon, evening) for 4 weeks (28 days) without any additional treatments for fatigue improvement. The control group participants consumed three 500 mL/bottle bottles of ordinary water (0 ppb of vanadium) three times a day for the same period of time without any additional treatments for fatigue improvement. After 4 weeks of VW consumption, total score of Revised Piper Fatigue Scale (RPFS) was reduced 30% compared to the basal score after Fisher's Exact Test which resulted in a statistical borderline significance (p=0.0799) and result of t-test of comparison between two groups' RPFS total score and fatigue cognitive/mood changes resulted in a statistical significance (p=0.0112). In detail, only 36.4% of control group (N=11) showed improvement while 77.8% of treatment group (N=9) showed fatigue improvement. No other lab measures, including thyroid hormone level indicated any significant differences between two groups. In conclusion, daily consumption of 1.5 L of vanadium water for 4 weeks showed improvement of non-advanced cancer patients' fatigue.

Association of CAST Gene Polymorphisms with Carcass and Meat Quality Traits in Chinese Commercial Cattle Herds

  • Li, Jiao;Zhang, Lu-Pei;Gan, Qian-Fu;Li, Jun-Ya;Gao, Hui-Jiang;Yuan, Zheng-Rong;Gao, Xue;Chen, Jin-Bao;Xu, Shang-Zhong
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.11
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    • pp.1405-1411
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    • 2010
  • Calpastatin (CAST), an endogenous inhibitor of the calpains, plays an important role in post-mortem tenderization of meat. The objectives of this study were to investigate single nucleotide polymorphisms (SNPs) in the bovine CAST gene and association with carcass and meat quality traits. A total of 212 cattle from commercial herds were tested in this study including 2 pure introduced breeds, 4 cross populations, and 3 pure Chinese native breeds. Five SNPs were identified at position 2959 (A/G), 2870 (G/A), 3088 (C/T), 3029 (G/A) and 2857 (C/T) in the CAST gene (GenBank Accession No. AF159246). Allele frequencies of SNP2959 and SNP2870 were 0.701 (A) and 0.462 (A), respectively. A general linear model was used to evaluate the associations between the two markers and 7 traits. The results showed that both SNP2959 and SNP2870 were significantly (p<0.01) associated with the Warner-Bratzler shear force (WBSF), while they had no significant association with the other 6 traits in the whole population. However, in Chinese native pure breeds, only SNP2870 had significant association with WBSF (p<0.05). The simultaneous analysis of two-marker genotype effects indicated animals containing the A/G haplotype (A for SNP2959 and G for SNP2870) tended to have lower shear force than those containing the G/A haplotype, and, especially, animals homozygous for the A/G haplotype had approximately 2 kg lower shear force than those homozygous for the G/A haplotype (p<0.01). These results suggested that both markers may be effective for the marker-assisted selection of meat quality traits in Chinese commercial herds, especially SNP2870 which can be used for Chinese native cattle.

An Antiviral Mechanism Investigated with Ribavirin as an RNA Virus Mutagen for Foot-and-mouth Disease Virus

  • Gu, Chao-Jiang;Zheng, Cong-Yi;Zhang, Qian;Shi, Li-Li;Li, Yong;Qu, San-Fu
    • BMB Reports
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    • v.39 no.1
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    • pp.9-15
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    • 2006
  • To prove whether error catastrophe /lethal mutagenesis is the primary antiviral mechanism of action of ribavirin against foot-and-mouth disease virus (FMDV). Ribavirin passage experiments were performed and supernatants of $Rp_1$ to $Rp_5$ were harvested. Morphological alterations as well as the levels of viral RNAs, proteins, and infectious particles in the BHK-21 cells infected using the supernatants of $Rp_1$ to $Rp_5$ and control were measured by microscope, real-time RT-PCR, western-blotting and plaque assays, respectively. The mutation frequency was measured by sequencing the complete P1- and 3D-encoding region of FMDV after a single round of virus infection from ribavirin-treated or untreated FMDV-infected cells. Ribavirin treatment for FMDV caused dramatically inhibition of multiplication in cell cultures. The levels of viral RNAs, proteins, and infectious particles in the BHK-21 cells infected were more greatly reduced along with the passage from $Rp_1$ to $Rp_5$, moreover, nucleocapsid protein could not be detected and no recovery of infectious virus in the supernatant or detection of intracellular viral RNA was observed at the $Rp_5$-infected cells. A high mutation rate, giving rise to an 8-and 11-fold increase in mutagenesis and resulting in some amino acid substitutions, was found in viral RNA synthesized at a single round of virus infection in the presence of ribavirin of $1000\;{\mu}M$ and caused a 99.7% loss in viral infectivity in contrast with parallel untreated control virus. These results suggest that the antiviral molecular mechanism of ribavirin is based on the lethal mutagenesis/error catastrophe, that is, the ribavirin is not merely an antiviral reagent but also an effective mutagen.

Quantitative Changes of PR Proteins and Antioxidative Enzymes in Response to Glomus intraradices and Phytophthora capsici in Pepper (Capsicum annuum L.) Plants

  • Zheng, Hu-Zhe;Kim, Yong-Woong;Lee, Hyun-Jin;Park, Ro-Dong;Jung, Woo-Jin;Kim, Young-Cheol;Lee, Sang-Hyun;Kim, Tae-Hwan;Kim, Kil-Yong
    • Journal of Microbiology and Biotechnology
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    • v.14 no.3
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    • pp.553-562
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    • 2004
  • To investigate protective activity in pepper plants, which were pre-inoculated with arbuscular mycorrhizal (AM) fungi Glomus intra radices (Gi), against pathogenic strain Phytophthora capsici (Pc), pathogenesis-related (PR) proteins and antioxidant enzymes were examined. The growth of root and shoot was the highest in peppers inoculated with G. intraradices, compared with non-inoculated control plants and those challenged by the pathogen with and without mycorrhizae after nine days of infection. Mycorrhizal colonization rate was reduced by about 10% in pathogen-challenged plants, but disease pressure was reduced. The activities of PR proteins, $\beta$-1- 3-glucanase and chitinase, were increased in Pc-treated plants compared to Gi+Pc-treated plants in leaves, but those in roots were suppressed. Superoxide dismutase activity and $H_2O_2${/TEX> content in Gi+Pc and Pc-treated plants were gradually increased in leaves. However, those in roots continuously increased up to 5 days, and then decreased dramatically. Peroxidase activity in leaves and roots increased after P. capsici infection both in plants inoculated with or without G. intraradices. These results suggest that AM fungi, G. intra radices, potentially act as one of the protective agents against plant pathogens. Changes of PR proteins and antioxidative enzymes in mycorrhizae-inoculated pepper appear to be regulated differently in leaves and roots by pathogen infection.

Prediction and analysis of structural noise of a box girder using hybrid FE-SEA method

  • Luo, Wen-jun;Zhang, Zi-zheng;Wu, Bao-you;Xu, Chang-jie;Yang, Peng-qi
    • Structural Engineering and Mechanics
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    • v.75 no.4
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    • pp.507-518
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    • 2020
  • With the rapid development of rail transit, rail transit noise needs to be paid more and more attention. In order to accurately and effectively analyze the characteristics of low-frequency noise, a prediction model of vibration of box girder was established based on the hybrid FE-SEA method. When the train speed is 140 km/h, 200 km/h and 250 km/h, the vibration and noise of the box girder induced by the vertical wheel-rail interaction in the frequency range of 20-500 Hz are analyzed. Detailed analysis of the energy level, sound pressure contribution, modal analysis and vibration loss power of each slab at the operating speed of 140 km /h. The results show that: (1) When the train runs at a speed of 140km/h, the roof contributes more to the sound pressure at the far sound field point. Analyzing the frequency range from 20 to 500 Hz: The top plate plays a very important role in controlling sound pressure, contributing up to 70% of the sound pressure at peak frequencies. (2) When the train is traveling at various speeds, the maximum amplitude of structural vibration and noise generated by the viaduct occurs at 50 Hz. The vibration acceleration of the box beam at the far field point and near field point is mainly concentrated in the frequency range of 31.5-100 Hz, which is consistent with the dominant frequency band of wheel-rail force. Therefore, the main frequency of reducing the vibration and noise of the box beam is 31.5-100 Hz. (3) The vibration energy level and sound pressure level of the box bridge at different speeds are basically the same. The laws of vibration energy and sound pressure follow the rules below: web