3.Effects of Folic Acid on Human T Lymphoid Leukemia Cell Line CEM Cells
li-jun, CHEN ; tai-e, ZHANG ; hong-mei, WANG ; fei, GAO ; yan, GU
Journal of Applied Clinical Pediatrics 1992;0(05):-
Objective To investigate the effects of the folic acid on human T lymphoid leukemia cell line CEM cells. Methods 1. MTT method was used to detect the proliferation of CEM cells co- cultured with folic acid of different concentrations and time;2. E-xamine the changes of morphology by light microscopy with Giemsa stain;3. Detect the percentage of apoptosis and cell cycle distribution as well as the expression of the apoptosis protein(Bcl- 2,C- myc) by flow cytometry;4. Detect DNA fragments by Agaiose elec-trophoresis;5. Detect the influence of folic acid to the anticancer effects of methotrexatc(MTX) by MTT methods. Results 1 Folic acid could inhibit the proliferation of CEM cells, and the optimal inhibitive concentrations range from 0. 4 ? 10-4 ?g/L to 3. 0 ? 10 -4 ?g/L,the inhibition rate was about 30% - 40% ; 2. Co - cultured with folic acid at 24,48, 72 hours, examined by light microscopy with Giemsa stain, apoptosis cells were found in all study groups but the higher apoptosis rate was found co - cultured with folic acid at concentration of (0.4 - 3.0) ? 10-4?g/L;3.The highest apoptosis rate was 6. 19% found at the concentration of 3 ? 10-4 ?g/L, but the cell cycle distribution had no statistical difference with control group, the expression of apoptosis related protein Bcl - 2 and C-myc was decreased;4 DNA was extracted from CEM cells co - cultured with 0.4? 10 -4 ?g/L and 3 ? 10-4 ?g/L folic acid for 48 hours. UNA ladders were visible by agarose electrophoresis of DNA fragments; 5. Folic acid did not affect the antitumor effect of MTX at the concentration from 0 2? 10-4 ?g/L to 12.0?10-4 ?g/L. Conclusion Folic acid may suppress proliferation and induce apoptosis of CEM cell
4.Electroacupuncture therapy and corrosion of acupuncture needle.
Yan-Hong SUN ; Tang-Yi LIU ; Hua-Yuan YANG ; Ming GAO ; Yin-E HU ; Gang XU ; Wen-Chao TANG
Chinese Acupuncture & Moxibustion 2014;34(12):1238-1240
During the process of electroacupuncture (EA) therapy, whether there being a corrosive effect in ac- upuncture needles was observed. Acupuncture needles were inserted into a rabbit's acupoint to perform a 12-hour electrical stimulation with three types of common EA waveform; additionally two needles were put in 0.9% sodium chloride solution with 12-hour direct current. Afterwards, environmental scanning electron microscope was applied to detect the surface physical characteristics of acupuncture needles. As a result, after a 12-hour continued electri- cal stimulation with three types of common EA waveform in the rabbit, there was no corrosive effect in acupunc- ture needles; but the direct current could cause severe corrosion in acupuncture needles. It is believed that there is no corrosion effect on acupuncture needles in current EA treatment, and some accidents reported in literature may be related to quality of EA device or improper manipulation during the treatment.
Acupuncture Points
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Animals
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Corrosion
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Electroacupuncture
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instrumentation
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Male
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Needles
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adverse effects
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Rabbits
5.Idiopathic arterial calcification of infancy:report of a case.
Feng-lan GAO ; Chun-ling LIU ; Yin-po ZHANG ; Yan-xin ZHANG
Chinese Journal of Pathology 2013;42(11):771-772
Autopsy
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Female
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Heart Failure
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etiology
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pathology
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Humans
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Infant
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Myocardial Infarction
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etiology
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pathology
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Vascular Calcification
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complications
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pathology
6.Effect of low-dose methylprednisolone on serum TNF-α level in children with Mycoplasma pneumoniae pneumonia.
Jin-E HE ; Chun-Yan GAO ; Hong-Ri LI
Chinese Journal of Contemporary Pediatrics 2013;15(10):850-853
OBJECTIVETo investigate the effect of low-dose methylprednisolone on serum tumor necrosis factor alpha (TNF-α) level in children with Mycoplasma pneumoniae pneumonia (MPP).
METHODSA case-control study was conducted among 38 children with MPP who received treatment in the Affiliated Hospital of Yan'an University between January and December 2012, and who had not received glucocorticoids before hospitalization. They were randomly divided into methylprednisolone treatment (n=20) and conventional treatment groups (n=18). The methylprednisolone treatment group was administered with methylprednisolone (1 mg/kg·d) by intravenous drip for three days in addition to conventional treatment. Serum samples were collected from both groups before treatment and on days 4 and 7 of treatment. Twenty-five children who underwent physical examination in the healthcare clinic during the same period were randomly selected as a normal control group, and serum samples were collected on the same day that the physical examination was performed. Serum TNF-α levels in the three groups were measured using enzyme-linked immunosorbent assay.
RESULTSOn admission, the methylprednisolone treatment and conventional treatment groups had significantly higher serum TNF-α levels than the normal control group (P<0.01), but there was no significant difference between the methylprednisolone treatment and conventional treatment groups. On days 4 and 7 of treatment, the methylprednisolone treatment group had significantly lower serum TNF-α levels than the conventional treatment group (P<0.05; P<0.01). On day 7 of treatment, there was no significant difference in serum TNF-α level between the methylprednisolone treatment and normal control groups, but the conventional treatment group still had a significantly higher serum TNF-α level than the normal control group (P<0.01).
CONCLUSIONSLow-dose methylprednisolone can significantly decrease serum TNF-α level and inhibit inflammatory response in children with MPP, and may reduce damage caused by inflammatory response.
Case-Control Studies ; Child ; Child, Preschool ; Female ; Humans ; Male ; Methylprednisolone ; administration & dosage ; therapeutic use ; Pneumonia, Mycoplasma ; drug therapy ; immunology ; Tumor Necrosis Factor-alpha ; blood
7.Intranasal administration of temozolomide for brain-targeting delivery: therapeutic effect on glioma in rats.
Ying LI ; Yongliang GAO ; Gang LIU ; Xiudi ZHOU ; Yan WANG ; Yulin WANG ; Lin MA
Journal of Southern Medical University 2014;34(5):631-635
OBJECTIVETo study the therapeutic effect of intranasal administration of temozolomide (TMZ) for brain-targeting delivery in a rat model bearing orthotopic C6 glioma xenografts.
METHODSForty Wistar rat bearing brain C6 glioma xenograft were randomly divided into 4 groups and treated with physiological saline solution or with TMZ by intravenous injection, gavage or intranasal administration. The tumor size, rat survival time and pathological changes were observed in each group.
RESULTSMagnetic resonance imaging showed a significantly reduced volume of glioma in intranasal TMZ group compared with that in the control, intraveneous TMZ injection group and TMZ gavage groups (12.45∓2.49 mm(3) vs 60.16∓4.12, 33.17∓3.56, and 35.16∓4.36 mm(3), respectively, P<0.05). The median survival time of the C6 glioma-bearing rats was also significantly longer in intranasal TMZ group than in the other 3 groups (31.0 days vs 20, 19, and 21.5 days, respectively, P<0.05). In the glioma xenografts, PCNA expression was the lowest and tumor cell apoptosis rate the highest in intranasal TMZ group.
CONCLUSIONIntranasal TMZ administration can suppress the growth of C6 glioma in rats and may serve as an effective strategy for glioma treatment.
Administration, Intranasal ; Animals ; Antineoplastic Agents, Alkylating ; administration & dosage ; Apoptosis ; Brain Neoplasms ; drug therapy ; Cell Line, Tumor ; Dacarbazine ; administration & dosage ; analogs & derivatives ; Drug Delivery Systems ; Glioma ; drug therapy ; Magnetic Resonance Imaging ; Neoplasm Transplantation ; Rats ; Rats, Wistar
8.Progress in epigenetic study of clear cell renal cell carcinoma
Yong-Sheng GAO ; Shao-Chun YAN ; Xiao-E JIA ; Guo SHAO
Chinese Journal of Pathophysiology 2018;34(1):178-182
The epigenetic changes of clear cell renal cell carcinoma(ccRCC )are considered to be the main molecular mechanisms of its pathogenesis ,including DNA methylation ,histone modification ,microRNA change ,and so on.DNA methyltransferases(DNMTs)catalyze the occurrence of DNA methylation.DNA methylation changes are mani-fested in the overall low methylation of the genome and the high methylation of specific sites ,which were involved in the de-velopment of ccRCC by affecting the expression of tumor suppressor genes.Due to histone-modified enzyme involvement ,histone modification is shown as possible genetic reversal.MicroRNA plays an important role in the abnormal expression of ccRCC genes.With the studies of epigenetic mechanism and molecular pathology ,it is important to explore the mechanisms and to seek effective early diagnosis ,treatment and prognosis intervention of ccRCC.
9.Detection of HPV 58 and cloning of its E7 gene in cervical cancer.
Yan-e GAO ; Ju ZHANG ; Zhong-can CHEN ; Tian-bao SONG ; Yan ZHAO ; Ge-lin ZHANG ; Xiao-jun YAN
Chinese Journal of Oncology 2004;26(9):543-546
OBJECTIVETo detect HPV 58, a common type of human papillomavirus (HPV), clone and express its E7 gene from biopsy specimens of cervical cancer.
METHODSHPV 58 from 58 biopsy tissues of cervical cancer was detected by GP5+/GP6+ PCR followed by template-directed dye-terminator incorporation assay with fluorescence polarization detection (TDI-FP). HPV 58 E7 gene was amplified from one HPV 58-positive sample, and then cloned into pGEM-T Easy vector. The recombinant plasmid, HPV58E7-pGEM-T was confirmed by sequencing. Subsequently, E7 gene was cloned into prokaryotic expression vector pRSET-A. The constructed pRSET-58E7 plasmids were transfected into BL21(DE3) cells, and induced to express 58 E7 protein by IPTG.
RESULTSAmong the 58 biopsy tissues of cervical cancer, 10 were HPV 58-positive, accounting for 19.2% of 52 HPV-positive cases. HPV 58 E7 gene was amplified from one HPV 58-positive sample. The constructed plasmids were identified containing HPV58 E7 gene by restriction enzyme analysis and sequencing. SDS-PAGE analysis showed that HPV58 E7 His6 fusion protein of M(r) 16 x 10(3) was expressed by pRSET-58E7 after induction by IPTG. The fusion protein accounted for 30% of total bacterial proteins.
CONCLUSIONHPV 58 is not uncommon in Chinese women with cervical cancer in Shaanxi province. Constructed HPV58 E7 recombinant plasmids can be effectively expressed in E.coli, which may provide a tool in diagnosis and vaccine design for HPV of HPV58-associated tumors.
Adult ; Cloning, Molecular ; Escherichia coli ; metabolism ; Female ; Genes, Viral ; Humans ; Middle Aged ; Papillomaviridae ; genetics ; isolation & purification ; Papillomavirus E7 Proteins ; biosynthesis ; genetics ; Papillomavirus Infections ; complications ; genetics ; Plasmids ; Recombinant Proteins ; biosynthesis ; genetics ; Transformation, Genetic ; Uterine Cervical Neoplasms ; genetics ; virology
10.Significance of B cell activating factor of TNF family promoter polymorphisms in patients with immune thrombocytopenic purpura.
Jun-Qing LIU ; Lin-Hua YANG ; Xiu-Hua CHEN ; Yan-Hong TAN ; Xiu-E LIU ; Jian-Fang CHEN ; Li-Xian CHANG ; Yan GAO
Journal of Experimental Hematology 2010;18(3):690-693
The study was aimed to examine the B cell activating factor promoter polymorphism of the TNF family (BAFF)-871 C/T in patients with immune thrombocytopenic purpura (ITP) and to explore its correlation with ITP and the relationship between the blood platelet count of newly diagnosed patients with ITP and genotypes of BAFF-871 C/T polymorphisms. Alleles specific polymerase chain reaction (ASP-PCR) and agarose gel electrophoresis were used to identify polymorphisms -871 C/T of BAFF promotor in 133 ITP patients and 117 healthy controls, and determine the genotype of subjects. Meantime, the frequency of genotype and alleles were analyzed. The results indicated that out of 133 patients with ITP, 33.1% patients exhibited C/C, 42.1% patients were heterozygous C/T, and 24.8% patients were homozygous T/T. The corresponding frequencies in 117 healthy controls were 55.6% C/C, 33.3% C/T and 11.1% T/T. The allele frequency of T in ITP patients and healthy controls were 45.9% and 27.4% respectively. There was significant difference in the BAFF-871 C/T genotypic frequency between the ITP patients and healthy controls (p < 0.05). The allele frequency of T in ITP patients was higher than that in healthy controls. There was no significant difference in the blood platelet counts between the various genotype (p > 0.05). It is concluded that the polymorphism -871 C/T of BAFF promoter is correlated with the pathogenesis of ITP. However, there is no significant difference in blood platelet counts between the various genotype, so the polymorphism -871 C/T of BAFF promoter can not be referred as the analysis index for evaluating the severity of ITP.
Adolescent
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Adult
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Aged
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Aged, 80 and over
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Alleles
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B-Cell Activating Factor
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genetics
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Case-Control Studies
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Child
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Female
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Gene Frequency
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Genotype
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Humans
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Male
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Middle Aged
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Polymorphism, Genetic
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Promoter Regions, Genetic
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Purpura, Thrombocytopenic, Idiopathic
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genetics
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Young Adult