1.Application of SCAR molecular marker technology in identification of Monascus strains.
Hong-Mei DING ; Zhi-Shan DING ; Hai-Bo LI ; Ni-Pi CHEN
China Journal of Chinese Materia Medica 2008;33(4):359-362
OBJECTIVETo establish an effective way for rapid identification of Monascus strains based on DNA molecular marker.
METHODA random amplified polymorphic DNA (RAPD) marker named F421 in genomic DNA of Monascus F strain was observed during a comparison of DNA fingerprints derived from 10 cultivated strains of Monascus. F421 was cloned and sequenced. Comparing the sequence of F421 (GenBank accession number EF063107) with other relative sequences in the GenBank databases, no distinct comparability was found. A pair of sequence characterized amplified region (SCAR) primers were designed based on the sequence of the cloned fragment and tested for the specific detection of Monascus F.
RESULTThe results of polymerase chain reaction showed that only a 421bp segment of Monascus F strain was amplified compared with other 9 cultivated strains of Monascus. And the acquired SCAR marker of strain F could be used as a specific DNA fingerprint to identify Monascus strain F within one day.
CONCLUSIONSCAR molecular marker technology is an effective new way to identify Monascus strains more rapidly. And also is an assistant tool to identify Monascus strains more accurately when disagreements come out using traditional classification. It could be applied widely to the protection of germ plasm resources, classification and identification distinguishing false strains of pharmaceutical fungi.
Molecular Sequence Data ; Monascus ; classification ; genetics ; Polymerase Chain Reaction ; methods ; Random Amplified Polymorphic DNA Technique
2.Study function of endophytic fungus in parasitism process of mistletoe.
Zhi-Shan DING ; Fu-Sheng JIANG ; Bo JIN ; Li XU ; Ni-Pi CHEN ; Gui-Yuan LV
China Journal of Chinese Materia Medica 2008;33(11):1243-1246
OBJECTIVETo research the function of endophytes of mistletoe in parasitism process of mistletoe in Pterocarya stenoptera.
METHODEndophytes from eight different parts of the mistletoe were separated by explant culture, and further screened by different CMC plates culture and DNS method to get cellulase high productive strains. The distribution of the endophytic fungus parasitized in mistletoe were prepared and stained to demonstrate by histological section of the intumescentia part of the P. stenoptera.
RESULTThe histological section indicated that aboundent of hyphasma were distributed around the haustorium of the mistletoe. Eighty three strains of endophytic fungus were separated, 38 of them were able to degrade cellulose, 19 strains showed high cellulase activity and 10 of which were separated from the parasitic position.
CONCLUSIONEndophytic fungus of mistletoe can secrete cellulase and assist the haustorium of mistletoe to breakthrough the cell walls as well as intercellular space tissues of the P. stenoptera, thus, the endophytic fungus plays an important role in the parasitism process of mistletoe in P. stenoptera.
Cellulase ; metabolism ; Cellulose ; metabolism ; Fungi ; metabolism ; Juglandaceae ; Symbiosis ; Viscum ; cytology ; microbiology
3.Protoplasts isolation, purification and plant regeneration of Pinellia cordata.
Xian YANG ; Dan-Dan MA ; Fu-Sheng JIANG ; Ni-Pi CHEN ; Bin DING ; Li-Xia JIN ; Chao-Dong QIAN ; Zhi-Shan DING
China Journal of Chinese Materia Medica 2014;39(21):4211-4215
The main factors which affected the isolation, purification and cultivation of Pinellia cordata protoplasts from leaves were studied. The results indicated that the optimum enzyme solution for P. cordata leaves was 13% CPW + 1.0% Cellulose +0.1% Pectolase, at pH 6.0, temperature (25-28 degrees C ) for 4 h. The sucrose density gradient centrifugation was adopted to purificate the protoplasts collected, when 25% sucrose was used as mediator, centrifugating at 500 rpm for 10 min. When the protoplasts were shallow liquid and liquid-solid double layer cultured on the medium of MS + 0.5 mg x L(-1) 6-BA + 0.25 mg x L(-1) NAA + 13% mannitol at the density of 2.5 x 104 protoplasts/mL, or fed and nursed cultured at the density of 100-500 protoplasts/mL, cell division could be observed for 3 days; granular calli appeared for 30 days. Calli was proliferated on the medium of MS + 0.5 mg x L(-1) 6-BA + 0.25 mg x L(-1) NAA solidified by 0.55% agar, and differentiated and regenerated after 5-6 months. Plant generation of P. cordata is successfully established.
Cell Separation
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methods
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Culture Media
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Pinellia
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physiology
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Protoplasts
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physiology
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Regeneration
4.Progress in the role of ubiquitin-proteasome system in hepatitis B virus X protein-mediated HBV infection and hepatocarcinogenesis
Yubo PI ; Yanmeng CHEN ; Ni TANG
Chinese Journal of Microbiology and Immunology 2023;43(7):517-524
Chronic hepatitis B virus (HBV) infection is one of the leading causes of hepatocellular carcinoma (HCC). Ubiquitin-proteasome system (UPS) mediates the post-translational modification and degradation of protein in eukaryotic cells. Recent studies have revealed that UPS plays an important role in HBV infection and hepatocarcinogenesis. Targeting HBx to intervene in the progression of HBV infection or HBV-related HCC has become a research hotspot both domestically and internationally in recent years. This article reviewed the progress in HBx promoting HBV infection and hepatocarcinogenesis through UPS.
5.Asymptomatic patients of chronic obstructive pulmonary disease in China.
Ming LU ; Wan-zhen YAO ; Nan-shan ZHONG ; Yu-min ZHOU ; Chen WANG ; Ping CHEN ; Jian KANG ; Shao-guang HUANG ; Bao-yuan CHEN ; Chang-zheng WANG ; Dian-tao NI ; Xiao-ping WANG ; Da-li WANG ; Sheng-ming LIU ; Jia-chun LÜ ; Ning SHEN ; Yan-ling DING ; Pi-xin RAN
Chinese Medical Journal 2010;123(12):1494-1499
BACKGROUNDChronic obstructive pulmonary disease (COPD) has a variable natural history and not all individuals follow the same course. This study aimed to identify the prevalence and characteristics of asymptomatic COPD patients from a population-based survey in China.
METHODSA multistage cluster sampling strategy was used in a population from seven different provinces/cities. All residents (over 40 years old) were interviewed with a standardized questionnaire and spirometry. Post-bronchodilator forced expiratory volume in 1 second (FEV(1))/forced vital capacity (FVC) of less than 70% was defined as the diagnostic criterion of COPD. All COPD patients screened were divided into symptomatic group and asymptomatic group according to the presence or absence of chronic respiratory symptoms. Socio-demographic, personal and exposure variables were collected and analyzed.
RESULTSAmong the 1668 patients who were diagnosed with COPD from the 25 627 sampling subjects, 589 (35.3%) were asymptomatic. The age, sex, body mass index (BMI), rural and urban distributions, smoking habit and education levels were similar in the two groups. A total of 64.7% of the asymptomatic patients had no comorbidities. Cardiovascular diseases and lung cancer were more common among symptomatic COPD patients than asymptomatic group. Asymptomatic COPD group were less likely to present with poor ventilation in the kitchen, a family history of respiratory disease and recurrent childhood cough. Asymptomatic COPD patients had significantly higher FEV(1) (73.1% vs. 61.0%), FVC (91.9% vs. 82.0%), and a higher ratio of FEV(1)/FVC (62.9% vs. 58.7%) (all P < 0.001) than symptomatic group. More asymptomatic patients were underdiagnosed (91.9% vs. 54.3%, P < 0.001) than symptomatic patients.
CONCLUSIONSThis large population-based survey confirmed a high prevalence of asymptomatic COPD patients in China. More use of spirometry screening test may be important to the early detection of COPD.
Adult ; Aged ; Aged, 80 and over ; China ; epidemiology ; Educational Status ; Female ; Humans ; Male ; Middle Aged ; Pulmonary Disease, Chronic Obstructive ; diagnosis ; epidemiology ; Risk Factors ; Smoking ; Spirometry ; Surveys and Questionnaires