3.Effects of ginsenoside Rh2(GS-Rh2) on cell cycle of Eca-109 esophageal carcinoma cell line.
Li LI ; Feng-ying QI ; Jun-ru LIU ; Lian-fu ZUO
China Journal of Chinese Materia Medica 2005;30(20):1617-1621
OBJECTIVETo investigate the effects of ginsenoside Rh2 (GS-Rh2) on growth inhibition and cell cycle of Eca-109 esophageal carcinoma cell line in culture.
METHODThe effects of GS-Rh2 on cell growth inhibition was detected by MTT assay. Cell cycle was analyzed by flow cytometry (FCM). Cell morphology was observed by a light microscope after HE staining. The protein expression of cell cycle components (cyclinE, CDK2, p21WAF1) were examined by immunocytochemistry and Western blot. The mRNA expression were examined by semiquantitative RT-PCR.
RESULTGS-Rh2 inhibited the proliferation of Eca-109 cells in dose and time-dependent manners. The inhibition rate was about 50% after 1-day treatment with 20 microg x mL(-1) GS-Rh2 x 20 microg x mL(-1) GS-Rh2 induced the mature differentiation and morphological reversion. With increasing dose of GS-Rh2 treatment, the cell number of G0/G1 phase was increased, whereas it decreased at S and G2/M phase. There was significant difference between 10, 20 microg x mL(-1) GS-Rh2 groups and the corresponding group without GS-Rh2 treatement. After treating cells by 20 microg x mL(-1) GS-Rh2 for 1, 2, 3 days individually, the protein and mRNA expression of both cyclinE and CDK2 reduced, while the expression of p21WAF1 enhanced gradually.
CONCLUSIONGS-Rh2 could arrest Eca-109 cells at G0/G1 phase and induce cell differentiation tending to normal. Furthermore, GS-Rh2 had an effect on expression of cell cycle components (cyclinE, CDK2 and p21WAF1) to inhibit Eca-109 cell proliferation.
Carcinoma, Squamous Cell ; pathology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclin E ; biosynthesis ; genetics ; Cyclin-Dependent Kinase 2 ; biosynthesis ; genetics ; Cyclin-Dependent Kinase Inhibitor p21 ; biosynthesis ; genetics ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Esophageal Neoplasms ; metabolism ; pathology ; Ginsenosides ; administration & dosage ; isolation & purification ; pharmacology ; Humans ; Panax ; chemistry ; Plants, Medicinal ; chemistry ; RNA, Messenger ; biosynthesis ; genetics ; Time Factors
4.Flow cytometric detection and significance of four cyclins in esophageal cancer.
Yun DU ; Xiao-ling WANG ; Guo-xiang WU ; Yong-jun WANG ; Hui-chai YANG ; Lian-fu ZUO
Chinese Journal of Oncology 2004;26(10):612-614
OBJECTIVETo study the expression and significance of cyclin E, cyclin D1, CDK4 and p27 protein in esophageal squamous cell cancer (ESCC) and their correlation with tumor differentiation and lymph node metastasis.
METHODSExpressions of cyclin E, cyclin D1, CDK4 and p27 protein in 65 patients with ESCC were quantitatively detected by flow cytometry.
RESULTSThe expressions of cyclin E, cyclin D1, CDK4 in poorly-differentiated ESCC were higher than those in well-differentiated ESCC (P = 0.0275, 0.0001, 0.0174). The expression of p27 in poorly-differentiated ESCC was lower than that in well-differentiated ESCC (P = 0.0042). There was positive correlation between cyclin E and cyclin D1, cyclin D1 and CDK4, but negative correlation between cyclin D1 and p27. The expressions of all four proteins were not correlated with lymph node metastasis.
CONCLUSIONThe expressions of cyclin E, cyclin D1, CDK4 and p27 are closely related to tumor differentiation of ESCC. An imbalance between positive and negative control of cell cycling might be critical in the carcinogenesis of esophageal squamous cell cancer.
Adult ; Aged ; Carcinoma, Squamous Cell ; metabolism ; secondary ; Cell Differentiation ; Cyclin D1 ; metabolism ; Cyclin E ; metabolism ; Cyclin-Dependent Kinase 4 ; metabolism ; Cyclin-Dependent Kinase Inhibitor p27 ; metabolism ; Cyclins ; metabolism ; Esophageal Neoplasms ; metabolism ; pathology ; Female ; Flow Cytometry ; Humans ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged
5.Mutation Analysis of CIB2 196C>T 272T>C and 297C>G in Patients with Non-syndromic Hearing Impairment
Qi LI ; Jun-Dong HE ; Jin-Li WANG ; Yong-Qin ZHENG ; Rong-Xia ZUO ; Ya-Lian SA
Journal of Kunming Medical University 2018;39(5):7-10
Objective To investigate the gene mutations of calcium-and integrin-binding protein 2 (CIB2) 196C>T, 272T > C and 297C > G carried by students with non-syndromic hearing impairment from special educational schools in Yunnan Province. Methods The experimental group included 337 students with non-syndromic hearing impairment who failed to carry deafness gene with GJB2 (35 del G, 176_191 del 16,235delC, 299_300 del AT), GJB3 (C538T,G547A), mtDNA 12S rRNA (A1555G, C1494T), and SLC26A4 (IVS7_2A>G, A2168G) . The control group consisted with 150 healthy people. Genomic DNA was isolated from peripheral blood with EDTA anti-coagulate. The subject's DNA fragments including CIB2 196C>T, 272T > C and 297C> G were amplified by polymerase chain reaction (PCR), and subsequently analyzed by direct sequencing to identify deafness-associated mutations. Results Both in the experimental group and control group, we failed to find the mutation of CIB2 196C>T, 272T>C and 297C>G in all individuals. Conclusion Mutations in CIB2 gene 196C>T, 272T>C and 297C>G are not a frequent cause of non-syndromic hearing loss among deaf people in Yunnan province. It provided important information for deafness with formulating landscape of gene screening in this region.
6.Disseminated cryptococcosis caused by Cryptococcus neoformans a case report and review
SHAN Kun ; ZUO Hui-fen ; ZHENG Cui-ying ; ZHANG Ze-kun ; ZHAO Lian-chun ; HUANG Yin-qi ; WANG Peng ; ZHAO Zhen-jun ; ZHANG Li-jie
China Tropical Medicine 2022;22(11):1043-
Abstract: To analyze the clinical, therapeutic and laboratory characteristics of disseminated cryptococcosis caused by Cryptococcus neoformans invading the blood stream in patient with liver cirrhosis and splenectomy. A 30-year-old male underwent splenectomy plus pericardial devascularization due to "splenomegaly and hypersplenism" in March in 2016. The patient had intermittent fever after operation for many times, and successively accompanied with back pain, left lower limb abscess and right hip pain. The highest body temperature was 39 ℃. CT and MRI revealed the lung lesion and multiple bone destruction. During that period, the effect of antibiotics was not good. On April 19th, 2017, Gram's stain, India ink stain, API 32C, Vitek 2 Compact, ribosomal ITS and IGS sequence analysis were performed to identify the strain isolated from the pus and blood stream. The serum of the patient was detected for cryptococcal antigen. Antifungal susceptibility test was used to determine drug sensitivity and minimum inhibitory concentration (MIC). The Cryptococcus neoformans isolated from fresh pus specimen showed a prominent, thick capsule after India ink stain. The colonies isolated from pus and blood stream were identified Cryptococcus neoformans using API 32C, Vitek 2 Compact, and sequence analysis of rDNA ITS and IGS. Cryptococcal capsule antigen was positive. The minimal inhibitory concentrations of 5-Flucytosine, amphotericin B, fluconazole, itriconazole, voriconazole against the isolate were <4 μg/mL, <0.5 μg/mL, 4 μg/mL, ≤0.25 μg/mL, 0.125 μg/mL respectively. The patient was initially treated with intravenous amphotericin B and flucytosine. After anti-Cryptococcus treatment for two months, the patient clinically improved, and the lesions were reduced on a follow-up CT scan. The patient made a full functional recovery after treatment for six months. Cryptococcosis has hidden onset, atypical clinical symptoms and lack of specificity. Blood stream is the main channel for Cryptococcus to spread and involve many organs of the whole body, including skin, bone and so on. Therefore, early use of blood culture to monitor blood flow dissemination, actively removing the primary focus and cutting off the infection route in time and carrying out effective anti-Cryptococcus treatment are conducive to the patient's early recovery.
7.Effects of sterigmatocystin, deoxynivalenol and aflatoxin G1 on apoptosis of human peripheral blood lymphocytes in vitro.
Xu-Ming SUN ; Xiang-Hong ZHANG ; Hui-Yan WANG ; Wen-Jun CAO ; Xia YAN ; Lian-Fu ZUO ; Jun-Ling WANG ; Feng-Rong WANG
Biomedical and Environmental Sciences 2002;15(2):145-152
OBJECTIVETo explore the effects of Sterigmatocystin (ST), Deoxynivalenol (DON) and Aflatoxin G1 (AFG1) on apoptosis of human peripheral blood lymphocytes (HPBLs) in vitro and thus to further elucidate the putative roles of these three mycotoxins on human immunosystem.
METHODSThe effects of ST, DON and AFG1 on apoptosis of HPBLs were studied with cell culture, flow cytometric (FCM) DNA analysis and DNA agarose gel electrophoresis.
RESULTSDNA agarose gel electrophoresis results showed the characteristic "ladder" pattern of apoptosis in HPBLs treated with ST, DON and AFG1. Flow cytometric DNA analysis revealed that typical subdiploid peaks of apoptosis in DNA histogram could be seen in all groups treated with the three mycotoxins. Significant time-effect and dose-effect relationships were found between the apoptosis rates and treatment time as well as concentrations of the three mycotoxins.
CONCLUSIONST, DON and AFG1 can induce apoptosis of HPBLs in vitro and may have some negative effects on human immunosystem.
Aflatoxins ; pharmacology ; Apoptosis ; drug effects ; Dose-Response Relationship, Drug ; Electrophoresis, Agar Gel ; Flow Cytometry ; Food Contamination ; Humans ; Lymphocytes ; cytology ; Sterigmatocystin ; pharmacology ; Time Factors ; Trichothecenes ; pharmacology
8.A study on the association between tea consumption and stroke.
Zuo CHEN ; Ying LI ; Lian-Cheng ZHAO ; Bei-Fan ZHOU ; Jun YANG ; Zeng-Wu WANG ; Min GUO ; Yang-Feng WU
Chinese Journal of Epidemiology 2004;25(8):666-670
OBJECTIVEThe aim of the study was to investigate the association between tea consumption and stroke.
METHODSA cross sectional study was conducted in autumn, 1998. The subjects were from 12 provinces in China. 15 groups of populations were selected by cluster randomized sampling and each group had about 1000 persons, aged from 35 to 60 years old. 14 212 subjects had complete data for analysis. Data regarding tea drinking would include drinking status, dose and type of teas. Logistic regression was used to analyze the association between tea consumption and stroke.
RESULTSThere was a strong inverse correlation between tea drinking and stroke after adjusting other risk factors of stroke (P < 0.05). The odds ratio (OR) of stroke was 0.60 [95% confidence interval (CI): 0.42 - 0.85] for subjects who drank tea compared to those who did not. Increased amount of tea consumption per month was associated with decreased stroke prevalence. The association for tea consumption over 150 gram per month and stroke was statistically significant (P < 0.05) with an OR value of 0.56 (95% CI: 0.36 - 0.89). Analytical results indicated that the OR value was 0.24 (95% CI: 0.06 - 1.01) for black tea and other tea (P = 0.05). The OR value was 0.35 (95% CI: 0.18 - 0.72) for green tea (P < 0.01), and 0.75 (95% CI: 0.51 - 1.11) for jasmine tea (P > 0.05).
CONCLUSIONTea drinking was independently associated with prevalence of stroke which might play a role in the prevention of the disease.
Adult ; China ; epidemiology ; Female ; Humans ; Hypertension ; epidemiology ; Logistic Models ; Male ; Middle Aged ; Plant Extracts ; therapeutic use ; Prevalence ; Sampling Studies ; Stroke ; epidemiology ; prevention & control ; Surveys and Questionnaires ; Tea ; chemistry
9.Nerve Growth Factor Promotes Angiogenesis and Skeletal Muscle Fiber Remodeling in a Murine Model of Hindlimb Ischemia.
Yong-Peng DIAO ; Feng-Kui CUI ; Sheng YAN ; Zuo-Guan CHEN ; Li-Shan LIAN ; Li-Long GUO ; Yong-Jun LI
Chinese Medical Journal 2016;129(3):313-319
BACKGROUNDTherapeutic angiogenesis has been shown to promote blood vessel growth and improve tissue perfusion. Nerve growth factor (NGF) has been reported to play an important role in both physiological and pathological angiogenesis. This study aimed to investigate the effects of NGF on angiogenesis and skeletal muscle fiber remodeling in a murine model of hindlimb ischemia and study the relationship between NGF and vascular endothelial growth factor (VEGF) in angiogenesis.
METHODSTwenty-four mice were randomly allocated to normal control group (n = 6), blank control group (n = 6), VEGF gene transfection group (n = 6), and NGF gene transfection group (n = 6). The model of left hindlimb ischemia model was established by ligating the femoral artery. VEGF165plasmid (125 μg) and NGF plasmid (125 μg) was injected into the ischemic gastrocnemius of mice from VEGF group and NGF group, respectively. Left hindlimb function and ischemic damage were assessed with terminal points at 21th day postischemia induction. The gastrocnemius of four groups was tested by hematoxylin-eosin staining, proliferating cell nuclear antigen and CD34 immunohistochemistry staining, and myosin ATPase staining. NGF and VEGF protein expression was detected by enzyme-linked immunosorbent assay.
RESULTSOn the 21th day after surgery, the functional assessment score and skeletal muscle atrophy degree of VEGF group and NGF group were significantly lower than those of normal control group and blank control group. The endothelial cell proliferation index and the capillary density of VEGF group and NGF group were significantly increased compared with normal control group and blank control group (P < 0.05). The NGF and VEGF protein expression of NGF group showed a significant rise when compared with blank control group (P < 0.05). Similarly, the VEGF protein expression of VEGF group was significantly higher than that of blank control group (P < 0.05), but there was no significant difference of the NGF protein expression between VEGF group and blank control group (P > 0.05). The type I skeletal muscle fiber proportion in gastrocnemius of NGF group and VEGF group was significantly higher than that of blank control group (P < 0.05).
CONCLUSIONSNGF transfection can promote NGF and VEGF protein expression which not only can induce angiogenesis but also induce type I muscle fiber expression in ischemic limbs.
Animals ; Antigens, CD34 ; metabolism ; Female ; Hindlimb ; metabolism ; pathology ; Ischemia ; metabolism ; pathology ; Mice ; Muscle, Skeletal ; metabolism ; pathology ; Neovascularization, Physiologic ; genetics ; physiology ; Random Allocation ; Vascular Endothelial Growth Factor A ; genetics ; physiology
10.OATP1B1 in drug-drug interactions between traditional Chinese medicine Danshensu and rosuvastatin.
Jin-hua WEN ; Xiao-hua WEI ; Xiao-hua CHENG ; Rong ZUO ; Hong-wei PENG ; Yan-ni LÜ ; Jian ZHOU ; Xue-lian ZHENG ; Jun CAI ; Yu-qing XIONG ; Li CAO
Acta Pharmaceutica Sinica 2016;51(1):75-79
The study was designed to explore the drug-drug interactions mechanisms mediated by OATP1B1 between traditional Chinese medicine Danshensu and rosuvastatin. First, the changes of rosuvastatin pharmacokinetics were investigated in presence of Danshensu in rats. Then, the primary rat hepatocytes model was established to explore the effects of Danshensu on the uptake of rosuvastatin by hepatocytes. Finally, HEK293T cells with overexpression of OATP1B1*a and OATP1B1*5 were established using a lentiviral delivery system to explore the effects of Danshensu on the uptake of rosuvastatin. Rosuvastatin pharmacokinetic parameters of C(max0, AUCO(0-t), AUC(0-∞) were increased about 123%, 194% and 195%, by Danshensu in rats, while the CL z/F value was decreased by 60%. Uptake of rosuvastatin in the primary rat hepatocytes was decreased by 3.13%, 41.15% and 74.62%, respectively in the presence of 20, 40 and 80 μmol x L(-1) Danshensu. The IC50 parameters was (53.04 ± 2.43) μmol x L(-1). The inhibitory effect of Danshensu on OATP1B1 mediated transport of rosuvastatin was related to the OATP1B1 gene type. In OATP1B1*5-HEK293T mutant cells, transport of rosuvastatin were reduced by (39.11 ± 4.94)% and (63.61 ± 3.94)%, respectively, by Danshensu at 1 and 10 μmol x L(-1). While transport of rosuvastatin was reduced by (8.22 ± 2.40)% and (11.56 ± 3.04)% and in OATP1B1*1a cells, respectively. Danshensu significantly altered the pharmacokinetics of rosuvastatin in rats, which was related to competitive inhibition of transport by OATPJBI. Danshensu exhibited a significant activity in the inhibition of rosuvastatin transport by OATP1B1*5-HEK293T, but not by OATP1B1*1a, suggesting a dependence on OATP1B1 sequence.
Animals
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Drug Interactions
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Drugs, Chinese Herbal
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pharmacology
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HEK293 Cells
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Hepatocytes
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drug effects
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metabolism
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Humans
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Lactates
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pharmacology
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Organic Anion Transporters
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metabolism
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Rats
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Rosuvastatin Calcium
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pharmacology
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Solute Carrier Organic Anion Transporter Family Member 1b1