1.The expression and significance of DNMT1 gene in oral squamous cell carcinoma
Ke ZHOU ; Hong MA ; Xiaofeng DUAN
Journal of Practical Stomatology 2016;32(4):584-586
The expression of DNA methyltranferase 1 (DNMT1 )mRNA and protein in 20 controls of normal oral mucosa tissue and 43 cases of oral squamous cell carcinoma(OSCC)was detected by Real-Time PCR and immunohistochemical staining and Western blot respectively. DNMTl mRNA CT values in OSCC and the controls were 0.958 6 ±0.986 6 and 0.459 5 ±0.525 8 respectively(Z =-2.028,P <0.05), The positive expression of DNMT1 protein in OSCC and the controls was 87% and 25% respectively(P <0.05).DNMT1 may play a role in the development of OSCC.
2.Radiation induced expression of HMGB1 in the cells with different radiosensitivity
Hong KANG ; Hui YANG ; Shaobo KE ; Fuxiang ZHOU ; Yunfeng ZHOU
Chinese Journal of Radiological Medicine and Protection 2016;36(7):486-490
Objective To investigate the effect ot ionizing radiation (IR) on the expressions of HMGB1 in the radiation-sensitive and radiation-resistant human cervical cancer cells and to analysis the role of HMGB1 in the regulation of radiosensitivity.Methods Human cervical cancer cells HeLa and its radioresistant strain HeLaR cells were irradiated with different doses of X-rays.The cells were collected at different time points after irradiation.The expressions of protein and mRNA of HMGB1 were detected by Western blot and real-time quantitative PCR.Results At the protein level,the expression of HMGB1 in HeLaR cells was significantly reduced at 6-36 h after 2,5 and 10 Gy X-ray irradiation (t =3.574-9.754,P <0.05),and then it was recovered to the control level at 48 h after IR.On the contrary,the expression of HMGB1 in HeLa cells was significantly increased at 6,12,48 h after 2 Gy IR (t =3.945-4.864,P<0.05),at 6,36,48hafter5 GyIR (t=-2.875-3.295,P<0.05),and at 36,48 h after 10 Gy IR (t =-4.480,-4.517,P < 0.05).At mRNA level,the trend of HMGB1 expression alteration was consistent with that of protein expression.Conclusions The changes of HMGB1 expression can be differently induced by X-rays in the human cervical cancer radiation-sensitivity cells and radiation-resistant cells.HMGB1 may be involved in the radioresistance of human cervical cancer.
3.Experience of treatment of subacute encephalopathy induced by 1, 2-dichloroethane poisoning.
Yuan-lin ZHOU ; Wei-jun HONG ; Shao-fa KE
Chinese Journal of Industrial Hygiene and Occupational Diseases 2009;27(4):253-254
Acute Disease
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Adult
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Ethylene Dichlorides
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poisoning
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Female
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Humans
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Male
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Neurotoxicity Syndromes
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therapy
5.Study on seed quality test and quality standard of Pesudostellaria heterophylla.
Cheng-Hong XIAO ; Tao ZHOU ; Wei-Ke JIANG ; Min CHEN ; Hou-Xi XIONG ; Ming-Wu LIAO
China Journal of Chinese Materia Medica 2014;39(16):3042-3047
Referring to the rules for agricultural seed testing (GB /T 3543-1995) issued by China, the test of sampling, seed purity, weight per 1 000 seeds, seed moisture, seed viability and germination rate had been studied for screening seed quality test methods of Pesudostellaria heterophylla. The seed quality from different collection areas was measured. The results showed that at least 6.5 g seeds should be sampled and passed through 10-mesh sieve for purity analysis. The weight of 1 000 seeds was determined by using the 500-seed method. The phenotypic observation and size measurement were used for authenticity testing. The seed moisture was determined under the higher temperature (130 ± 2) degrees C for 5 hours. The seeds were dipped into 0.2% TTC sustaining 1 hour at 40 degrees C, then the viability could be determined. The break dormancy seeds were cultured on sand at 10 degrees C. K cluster analysis was applied for the data analysis, the seed quality from different collection areas grading of P. Heterophylla was described as three grades. The seed quality of each grade should reach following requirements: for first grade seeds, germination rate ≥ 86%, 1 000-grain weight ≥ 2.59 g, purity ≥ 87%, moisture ≤ 13.1%; for second grade seeds, germination rate ≥ 70%, 1 000-grain weight ≥ 2.40 g, purity ≥ 77%, moisture ≤ 14.3%; for third grade seeds, germination rate ≥ 41%, 1 000-grain weight ≥ 2.29 g, purity ≥ 76%, moisture ≤ 15.8%. The seed testing methods for quality items of P. heterophylla had been initially established, as well as the primary P. heterophylla seed quality classification standard.
Caryophyllaceae
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chemistry
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growth & development
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Germination
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Quality Control
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Seeds
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chemistry
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growth & development
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Water
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analysis
6.Case study on the pay by segmentation and quota in NRCMS
Heng WANG ; Dian ZHOU ; Beihai XIA ; Hong DING ; Zhaohua KE ; Zhigang HE
Chinese Journal of Hospital Administration 2009;25(4):277-280
Objective To refrain the medical costs from out-of-control increase, and identify a per-disease payment mode suitable for New-CMS.Methods Case studies were conducted on all the data of five diseases in the course of three years, in a field study of the pilot counties for NRCMS in Anhui Province.Results This system of per-disease "pay by segmentation and quota" is composed of five parts: choice of diseases, measurement of payment criteria, method of settlement, method of compensation, and methods of supervision.Conclusion This system is an effective way to keep the medical costs in the NRCMS under control, given an effective play of the five supportive measures including the clinical pathways for individual diseases.
7.Identification of Bletillae Rhizoma and its adulterants by SNPs in ITS2.
Dan ZHAO ; Tao ZHOU ; Wei-ke JIANG ; Cheng-hong XIAO ; Chuan-zhi KANG
China Journal of Chinese Materia Medica 2015;40(18):3573-3578
To establish a molecular identification method for Bletillae Rhizoma, this paper extracted genome DNA from Bletillae Rhizoma and its adulterants. The sequences of rDNA ITS2 were sequenced after amplifying. Then multiple alignments of ITS2 were constructed phylogenetic tree with Neighbor Joining by MEGA 5. 1 and found out SNPs loci. The result showed that rDNA ITS2 region could identify Bletillae Rhizoma and its adulterants. There existed the SNPs loci, which could identify Bletilla striata and B. ochracea. Furthermore, we designed specific primers against the SNPs loci of B. striata and B. ochracea, then screened primers and optimized the PCR amplification conditions. Finally, the DNA of B. striata and B. ochracea were specifically amplified by BJ59-412F, BJ59-412R and HHBJ-225R. The length of amplification products were respectively about 350 bp and 520 bp that were effectively identified of B. striata and B. ochracea. While, the adulterants of Bletillae Rhizoma were no-reaction occurring. To sum up, the amplification conditions of the primers can identify B. striata, B. ochracea and their adulterants successfully at the same time. This method was easy, time-saving, and reliable, which can be used as a rapid method for molecular identification of Bletillae Rhizoma.
Base Sequence
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DNA Primers
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genetics
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DNA, Intergenic
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genetics
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DNA, Plant
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genetics
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Drug Contamination
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prevention & control
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Molecular Sequence Data
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Orchidaceae
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classification
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genetics
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Phylogeny
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Polymorphism, Single Nucleotide
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Rhizome
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classification
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genetics
8.Rapid molecular identification of pseudostellariae radix.
Dan ZHAO ; Tao ZHOU ; Wei-Ke JIANG ; Yuan YUAN ; Cheng-Hong XIAO ; Wei ZHENG
China Journal of Chinese Materia Medica 2014;39(19):3689-3694
To establish a convenient and rapid method for identification of Pseudostellariae Radix by molecular identification, the rDNA-ITS sequences of Pseudostella riaheterophylla and its adulterants had been aligned to find out specific fragment. The specific primers against the fragment were designed and the PCR amplification conditions were optimized. The fluorescence reaction of the PCR products colored by 100 x SYBR Green I was observed under UV. The concentration of reaction buffer included 5.5 μL DNA Taq polymerase premix, 10 pmol Tzs-2F and 10 pmol Tzs-2R, 20-80 ng template DNA, and plus double sterile distilled water to 25 μL. The PCR thermal profile was as follows: predenaturation at 95 degrees C for 1 min, followed by 30 cycles of denaturation at 95 degrees C for 5 seconds, primer annealing and extension at 56 degrees C for 15 seconds, then it was extension at 72 degrees C for 30 seconds. The fluorescence reaction of Pseudostellariae Radix showed green fluorescence, while adulterants had not. Extraction, amplification DNA and all steps of molecular identification could be completed successfully in 40 minutes. The approach could amplify DNA template of Pseudostellariae Radix specificity, and its product with 1 μL 100 x SYBR Green I could engender green fluorescence under UV. The method was simple and accurate, so it could be used for identification of Chinese traditional medicine.
Caryophyllaceae
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classification
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genetics
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DNA Primers
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genetics
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Drug Contamination
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prevention & control
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Plant Roots
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classification
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genetics
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Polymerase Chain Reaction
;
methods
9.Effect of peroxisome proliferator-activated receptor-gamma excitomotor on the expression of nuclear factor-kappa B and apoptosis of retinal ganglion cells in rat retina with diabetes mellitus
Wen-jun, GOU ; Ke, OU-YANG ; Hong-bin, LV ; Qing-lan, LI ; Qi, ZHOU ; Jun, ZHANG
Chinese Journal of Experimental Ophthalmology 2012;30(8):709-714
Background As one of the most common microvascular complication of diabetes in eyes,diabetic retinopathy (DR) is one of the most important cause of blindness.Nuclear factor-kappa B (NF-κB) is involved in the occurrence and development of the disease through the activation of a series of inflammatory cytokines.Objective The present study was to investigate the effects of peroxisome proliferator-activated receptor-gamma (PPAR-γ) excitomotor,rosiglitazone,on NF-κB expression and apoptosis of the retinal ganglion cells (RGCs) in the retina with diabetes mellitus. Methods Ninety SPF male Wistar rats were randomized into normal control group,diabetic control group and rosiglitazone group.Diabetes mellitus was induced by intraperitoneal injection of 50 mg/kg streptozotocin(STZ).Then 3 mg/kg rosiglitazone was intragastricly administered once per day in the rosiglitazonegroup,and the same volume of saline solution was used at the same way in the normal control group and diabetic control group from 3 days after modeling.The rats were sacrificed and the eye cups specimen was made at 4,8 and 12 weeks after usage of drugs.Retinal histopathological examination was performed by hematine-eosin staining,and expression of NF-κB p65 protein in retina and apoptotic index(AI) of RGCs were detected by immunohistochemistry and TUNEL assay,respectively in different time points mentioned above.The use of the animals complied with the Regulations for the Administration of Affairs Concerning Experimental Animals by State and Technology Commission.Results The blood glucose level was significantly elevated at various time points in the diabetic control group and rosiglitazone group compared with normal control group (P<0.01 ),and that of the rosiglitazone group was significantly declined in comparison to the diabetic control group (q =0.81,0.82,1.23,P> 0.05 ).Normal retinal structure was seen in the normal control group,and edema retinal cell and disorder of retinal layers were exhibited in the diabetic control group.Retinal structure was almost normal in the rosiglitazone group.The NF-κB p65 was expressed weakly in the retina of normal control group,but the expression of NF-κB p65 was significantly elevated in the diabetic control group and rosiglitazone group compared with the normal control group(P<0.01 ).However,the expression of NF-κB p65(A value)was significantly decreased in the rosiglitazone group compared with diabetic control group at 8 weeks and 12 weeks( q=17.77,15.30,P<0.01 ).There were a few apoptotic cells in rat retina of the normal control group.Compared with the normal control group,the AI of the diabetic control group and rosiglitazone group was significantly reduced(P<0.01 ).However,the AI of RGCs in the rosiglitazone group was significantly lower than that of diabetic control group in various time points (q =19.28,27.39,49.92,P<0.01 ). Conclusions As one of the PPAR-γexcitomotors,rosiglitazone can inhibit apoptosis of RGCs through downregulating the expression of NF-κB in rat retina with diabetes mellitus,indicating a protective effect of rosiglitazone on retina in diabetic rat.
10.Radiosensitization of clioquinol and zinc in human cervical cancer cell lines
Shan LU ; Yuan KE ; You WANG ; Hong ZHAO ; Xiaojia GAO ; Haijun YU ; Fuxiang ZHOU
Chinese Journal of Radiological Medicine and Protection 2016;36(4):241-245
Objective To investigate the radiosensitization effects of the combination treatment of clioquinol (CQ) and zinc on human cervical cell line HeLa in vitro.Methods Cells were divided into the 4 groups:controls,drug,radiation,and combined drug and radiation group.Cytotoxic effect of CQ and zinc on cell viability was determined by CCK-8 assay.Radiosensitization effect of CQ and zinc on HeLa cells was detected by colongenic assay,and the single-hit multi-target model was used to stimulate the doseresponse curve of survival and to calculate radiosensitization parameters.The cell cycle and apoptosis of HeLa cells were analyzed with flow cytometry.Luciferase reporter assay was used to study NF-κB activity of HeLa cells.Results The combination of CQ and zinc inhibited cell growth in a dose-dependent manner (F =188.00,P < 0.01).The mean lethal dose was 3.16 and 2.04 Gy for radiation group and combined drug and radiation group,respectively,and hence the SER was 1.55.Compared with the radiation group,the ratio of G2-phase cells in the combined drug and radiation group decreased(t =10.39,P < 0.05),the apoptosis rate increased at 24 h post-irradiation (t =5.64,P < 0.01),and the NF-κB activity decreased (t =21.42,P < 0.05).Compared to the control group,the NF-κB activity increased in the radiation group(t=6.23,P<0.05),but decreased in the drug group(t =12.48,P<0.05).Conclusions The combination of CQ and zinc could increase the radiosensitivity of HeLa cells by decreasing the ratio of G2-phase cells,increasing apoptosis and the inhibiting of NF-κB activity.