1.Antitumor activity of recombinant antimicrobial peptide penaeidin-2 against kidney cancer cells.
Ming-Xiang, MENG ; Jian-Fang, NING ; Jing-You, YU ; Dan-Dan, CHEN ; Xiao-Lin, MENG ; Jin-Ping, XU ; Jie, ZHANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2014;34(4):529-34
Penaeidin-2 (Pen-2) is an important antimicrobial peptide derived from the Pacific white shrimp, Penaeus vannamei, and possesses both antibacterial and antifungal activities. Recent studies suggest that recombinant penaeidins show similar activities to the native Pen-2 protein. Previous researches have shown that some antimicrobial peptides (AMPs) exhibit cytotoxic activity against cancer cells. To date, there have been no studies on the antitumor effects of Pen-2. This study evaluated the potential of recombinant pen-2 (rPen-2) in the selective killing of kidney cancer cell lines ACHN and A498, and its action mechanism. MTT assays found the maximal growth inhibition of HK-2, ACHN and A498 cells treated with 100 μg/mL rPen-2 at 48 h was 13.2%, 62.4%, and 70.4%, respectively. DNA-specific fluorescent dye staining showed a high percentage of apoptosis on cancer cells. Flow cytometry revealed that the apoptosis rate of HK-2, ACHN and A498 cells was 15.2%, 55.2%, and 61.5% at 48 h respectively, suggesting that rPen-2 induced higher apoptosis rate in cancer cells than in HK-2 cells. Laser confocal scanning microscopy demonstrated that the plasma membrane was the key site where rPen-2 interacted with and destroyed tumor cells. Scanning electron microscopy showed the morphologic changes of the cell membranes of kidney cancer cells treated with rPen-2. These results suggest that rPen-2 is a novel potential therapeutic agent that may be useful in treating kidney cancers.
2.Role of Bcl-2 signal pathway in apigenin preconditioning against cardiomyocytes anoxia/reoxygenation injury
Min ZHOU ; Jiazhen YOU ; Huan HE ; Dan LIU ; Zhangping LIAO ; Lei TANG ; Dong YIN ; Ming HE
Chinese Pharmacological Bulletin 2015;(1):122-126,127
Aim To investigate the relationship be-tween the cardioprotection of apigenin ( Api ) from an-oxia/reoxygenation ( A/R) injury and Bcl-2 pathway. Methods H9 c2 cardiomyocytes were cultured and di-vided into normal control group, A/R group, Api pre-treatment group ( Api ) , Api + Bcl-2 inhibitor group ( Api + ABT-737 ) . Expression of Bcl-2 was deter-mined by Western blot,and cell viability was measured by MTT method. LDH, SOD, GSH-Px, MDA activity were determined by chromometry. ROS generation, mi-tochondrial membrane potential and apoptosis were de-termined by flow cytometry. Results 25h after apige-nin precondition,the expression of Bcl-2 was upregulat-ed in cardiomyocytes ( P <0. 01 ) . In the group pre-treated with 40 μmol · L-1 apigenin before A/R, the activity of LDH in culture medium decreased; the ac-tivity of intracellular SOD, GSH-Px increased; the content of MDA and ROS generation decreased; cell viability increased; mitochondrial membrane potential could be more stable and cell apoptosis decreased ( P<0. 01 ) . However, all these protective effects were attenuated significantly in the group pretreated with apigenin and Bcl-2 inhibitor ABT-737 . Conclusion The effect of apigenin against A/R injury in cardiomyo-cytes involves Bcl-2 pathway, and at least partly de-pends on its effect on upregulating the expression of Bcl-2 .
3.Study on acting mechanism of shenqi yiqi drop pill for intervening irido-microangiopathy in diabetic rats.
Ming JIN ; Hai-dan LIU ; You-hua ZHANG
Chinese Journal of Integrated Traditional and Western Medicine 2010;30(2):174-177
OBJECTIVEUse laser confocal microscopy overspeed camera technique and fluorescence albumin labeling to study the acting mechanism of Qishen Yiqi Drop Pill (QYDP) for intervening irido-microangiopathy (IMAP) in diabetic rats.
METHODSRat model of diabetes mellitus type 1 (DM1) was established by intraperitoneal injection of streptozocin (STZ). The model rats were randomly divided into three groups, the treatment group, the model group and the control group. At the same time a normal control group was set up. The treatment group was medicated with QYDP (prepared into liquid form), and the control group with Duobeisi liquor (1 g/kg per day) for 10 months. The dynamic state of iris microcirculation in rats was observed using laser confocal microscopy overspeed camera.
RESULTSCompared with the treatment group, blood flow in iris of model rats was slower significantly (P < 0.01); the fluorescence density and leakage area of inside and outside iris vessels, and the iris vascular diameter were significantly higher in the model group than those in the treatment group (P < 0.01).
CONCLUSIONQYDP has definite effect in improving iris microcirculation, which can accelerate the blood flow, inhibit the abnormal expansion of vessels and improve the increased iris micro-vascular permeability.
Animals ; Capillary Permeability ; drug effects ; Diabetes Mellitus, Experimental ; complications ; drug therapy ; Diabetes Mellitus, Type 1 ; complications ; drug therapy ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Iris ; blood supply ; Iris Diseases ; etiology ; prevention & control ; Male ; Phytotherapy ; Rats ; Rats, Sprague-Dawley
5.Association between vitamin D receptor gene polymorphisms and pediatric Crohn's disease in China: a study based on gene sequencing.
You-You LUO ; Xiao-Li SHU ; Hong ZHAO ; Jin-Dan YU ; Ming MA ; Jie CHEN
Chinese Journal of Contemporary Pediatrics 2013;15(11):1006-1008
OBJECTIVETo investigate the association between TaqI, BsmI, and ApaI polymorphisms of vitamin D receptor (VDR) gene and pediatric Crohn's disease (CD) in China.
METHODSNineteen children with CD were selected as a case group, and 122 healthy children who underwent physical examination were selected as a control group. Serum 25-hydroxyvitamin D3 [25(OH)D3] levels were measured using ELISA. The TaqI, BsmI, and ApaI polymorphisms of VDR gene were determined by gene sequencing, and the two groups were compared in terms of genotype and allele frequencies.
RESULTSThe case group had significantly lower serum 25(OH)D3 levels than the control group (17.3±2.4 ng/mL vs 26.9±2.1 ng/mL; P<0.05). There were no significant differences in the frequencies of genotypes and alleles of TaqI, BsmI, and ApaI polymorphisms between the case and control groups (P>0.05).
CONCLUSIONSChildren with CD have low serum 25(OH)D3 levels. TaqI, BsmI, and ApaI polymorphisms of VDR gene may not be associated with susceptibility to CD among the Chinese population.
Adolescent ; Calcifediol ; blood ; Child ; Child, Preschool ; Crohn Disease ; blood ; genetics ; Female ; Humans ; Male ; Polymorphism, Single Nucleotide ; Receptors, Calcitriol ; genetics ; Sequence Analysis, DNA
6.Roles of UGT 1A1 gene mutation in the development of neonatal hyperbilirubinemia in Guangxi.
Zong-yan GAO ; Dan-ni ZHONG ; Yi LIU ; You-nan LIU ; Lu-ming WEI
Chinese Journal of Pediatrics 2010;48(9):646-649
OBJECTIVENeonatal unconjugated hyperbilirubinemia is one of the most common conditions encountered by the practicing pediatricians. Although it is usually self-limited and benign, the condition is of importance because of the rare instances in which severe hyperbilirubinemia can lead to bilirubin encephalopathy or kernicterus. The uridine diphosphate-glucuronosyl transferase 1A1 (UGT 1A1) gene controls bilirubin conjugation by determining the structure of the enzyme glucuronosyltransferase, which is synthesized in the hepatocyte. In the recent years much has been learned about the relationship between UGT 1A1 gene mutation and neonatal hyperbilirubinemia. This study aimed to investigate the roles of UGT 1A1 gene mutation in the development of neonatal hyperbilirubinemia in Guangxi.
METHODSA total of 73 cases with hyperbilirubinemia and 31 healthy neonates were enrolled. UGT 1A1 G71R genotypes were identified by the (amplification refractory mutation system, ARMS) and direct sequencing method in all the neonates. To analyze the incidence of bilirubin encephalopathy, the peak (total serum bilirubin, TSB) concentration after 72 hours of age, and the possibility of TSB > 20 mg/dl of each group.
RESULTS(1) The frequencies of allele G71R were 0.1915 in this study, 0.2329 in hyperbilirubinemia group vs. 0.097 in healthy groups. The allele gene frequency of G71R in neonatal hyperbilirubinemia was higher than that in the normal group (P < 0.05). (2) Homozygous neonates had higher possibility to develop bilirubin encephalopathy and higher TSB concentration 72 hours after birth (28.57%, 23.12 ± 4.58) than the normal group (0%, 17.68 ± 2.69). The difference between the former two was significant (P < 0.001). (3) The TSB of the 5 neonates was > 20 mg/dl in G71R homozygous type, the odds ratio and 95%CI were 7.955 (1.349, 46.899).
CONCLUSION(1) G71R mutation gene was associated with neonatal jaundice in Guangxi region. (2) The possibility of TSB > 20 mg/dl in G71R homozygous was higher than those of the wild-type. (3) The incidence of bilirubin encephalopathy and TSB concentration after 72 hours of age for neonates who were homozygous to G71R gene were higher than the wild-type.
Case-Control Studies ; China ; epidemiology ; Genotype ; Glucuronosyltransferase ; genetics ; Humans ; Hyperbilirubinemia, Neonatal ; epidemiology ; genetics ; Infant, Newborn ; Mutation
7.Pharmacokinetics of the combined preparation of lisinopril and hydrochlorothiazide on Chinese healthy volunteers.
Yang WEI ; Ying-Ying SUN ; Ke-Shen WANG ; Dan-Li SUN ; Ming-You ZHENG
Acta Pharmaceutica Sinica 2011;46(8):955-961
The aim of the present study, performed on two different groups of volunteers, is to characterize the pharmacokinetics of lisinopril/hydrochlorothiazide combined tablet. After administration of high, medium and low doses of lisinopril/hydrochlorothiazide combined tablets, AUC and C(max) of two compounds both increase significantly with increase of dose. Neither normalized AUC/Dose nor C(max)/Dose has significant difference between every two tested dose groups. The similar results can be observed as for the parameters of t(max). Lisinopril and hydrochlorothiazide are both eliminated with linear characteristics. After repeated administration of lisinopril/hydrochlorothiazide combined tablets, AUC, C(max) and C(min) of lisinopril in the steady state increase. AUC and C(min) increase significantly. As for hydrochlorothiazide, AUC, C(max), C(min), and t(max) also increase in steady state. AUC and C(min) increase significantly. Administered with the test medication, lisinopril has an fluctuation index (FI) value of 2.29 and reaches a relative steady concentration. But hydrochlorothiazide has an FI value of 4.09 with relatively large fluctuating concentrations.
Adult
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Antihypertensive Agents
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administration & dosage
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blood
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pharmacokinetics
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Area Under Curve
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Asian Continental Ancestry Group
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Biological Availability
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Cross-Over Studies
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Dose-Response Relationship, Drug
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Drug Combinations
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Female
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Humans
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Hydrochlorothiazide
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administration & dosage
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blood
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pharmacokinetics
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Lisinopril
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administration & dosage
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blood
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pharmacokinetics
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Male
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Tablets
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Young Adult
8.Relationship between glucose-6-phosphate dehydrogenase gene mutations and neonatal jaundice in Naning, Guangxi.
Dan-Ni ZHONG ; Zong-Yan GAO ; You-Nan LIU ; Yi LIU ; Lu-Ming WEI
Chinese Journal of Contemporary Pediatrics 2009;11(12):970-972
OBJECTIVETo study the correlation between glucose-6-phosphate dehydrogenase (G-6-PD) activities and three common mutations of G-6-PD gene G1388A, G1376T and A95G and investigate the effects of G-6-PD gene mutations on neonatal jaundice in Nanning, Guangxi.
METHODSOne hundred and twenty-four neonates from Nanning, Guangxi, with hyperbilirubinemia were enrolled. The ARMS-PCR and PCR/REA methods were used to determine G-6-PD gene mutations. G-6-PD activities were measured using the NBT method. The incidence of acute bilirubin encephalopathy and the peak bilirubin concentration 72 hrs after birth were compared between the neonates with different genotypes and between the G-6-PD mutation and normal groups. The risk of blood serum bilirubin >340 mumol/L was evaluated by logistic regression analysis.
RESULTSOf the 124 cases, gene mutations were found in 37 cases, including G1388A (n=20), G1376T (n=14), A95G (n=4) and G1388A+A95G (n=1). Five cases (25%) showed normal G-6-PD activities in the G1388A gene mutation group and 4 (29%) had normal G-6-PD activities in the G1376T G1388A gene mutation group. All of 4 cases of A95G G1388A gene mutation showed a deficiency of G-6-PD activities. There were no significant differences in the incidence of acute bilirubin encephalopathy and the peak bilirubin concentration 72 hrs after birth between the G1388A and G1376T G1388A gene mutation groups. The incidence of acute bilirubin encephalopathy, the peak bilirubin concentration 72 hrs after birth and the risk of serum bilirubin >340 micromol/L in the G-6-PD mutation group were not different from the normal group.
CONCLUSIONSG1388A, G1376T and A95G are common G-6-PD gene mutations in Nanning, Guangxi. The false negative results may be received when the NBT method is used for diagnosis of G-6-PD deficiency. There are similar effects on the incidence of acute bilirubin encephalopathy and the peak bilirubin concentration 72 hrs after birth between different gene mutation groups. G-6-PD gene mutations alone may not contribute to the development of acute bilirubin encephalopathy and the changes of peak bilirubin concentration 72 hrs after birth and the risk of serum bilirubin >340 micromol/L.
Bilirubin ; blood ; Encephalitis ; etiology ; Female ; Genotype ; Glucosephosphate Dehydrogenase ; genetics ; metabolism ; Glucosephosphate Dehydrogenase Deficiency ; genetics ; Humans ; Infant, Newborn ; Jaundice, Neonatal ; blood ; genetics ; Male ; Mutation
9.Investigation on dietary nutrients among rural stranded children of 2-7 year olds in China
Dan-Hui DUAN ; Ming-Yuan ZHU ; Jia-You LUO ; Zheng WANG ; Chu-Hui FU ; Wei-Min ZHANG ; Mei-Ling YAO ; Gui-Qin DUAN
Chinese Journal of Epidemiology 2009;30(4):326-330
Objective To understand the dietary nutrients among rural stranded children.Methods 2551 children aged 2 to 7, including 1278 stranded children in the rural areas and another 1273 children served as controls were selected, using multistage stratified cluster random sampling. Dietary survey was performed with three-day weighing dietary method and questionnaire on food intake. Data on diet were analyzed and evaluated by the Dietary Reference Intakes (DRIs) recommend by Chinese Nutrition Society, to evaluate the levels on energy and nutrient intake among stranded children in the rural areas. Results The dietary pattern among rural stranded children mainly consisted of grains and vegetables, but the intakes of animal products, fruits, and snacks were significantly less than in the control group. The intakes of three major energy-producing nutrients were below the recommend nutrient intake. Minerals as calcium, zinc, selenium, kalium and vitamins as vitamin A, B1, B2were insufficient.Most of the rural stranded children took nutrients insufficiently, with 50% lack of adequate energy and 80% of protein, 90% of minerals (calcium, zinc etc.) and vitamins (vitamin B1 and vitamin B2 etc.).Sources from high quality protein was insufficient, only consisting 35% of the total protein, but overabundant (over 64 % ) from the plants. The intake of plant-sourced iron was overabundant, accounted for 87%. Conclusion The dietary pattern was unsatisfactory with insufficient intake of energy-sourced proteins and some nutrients. The sources of energy, protein, and iron were mostly obtained from underbalaneed foods. It is necessary to improve the dietary nutrients status among rural stranded children aged 2-7 years.
10.Antitumor activity of recombinant antimicrobial peptide penaeidin-2 against kidney cancer cells.
Ming-xiang MENG ; Jian-fang NING ; Jing-you YU ; Dan-dan CHEN ; Xiao-lin MENG ; Jin-ping XU ; Jie ZHANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2014;34(4):529-534
Penaeidin-2 (Pen-2) is an important antimicrobial peptide derived from the Pacific white shrimp, Penaeus vannamei, and possesses both antibacterial and antifungal activities. Recent studies suggest that recombinant penaeidins show similar activities to the native Pen-2 protein. Previous researches have shown that some antimicrobial peptides (AMPs) exhibit cytotoxic activity against cancer cells. To date, there have been no studies on the antitumor effects of Pen-2. This study evaluated the potential of recombinant pen-2 (rPen-2) in the selective killing of kidney cancer cell lines ACHN and A498, and its action mechanism. MTT assays found the maximal growth inhibition of HK-2, ACHN and A498 cells treated with 100 μg/mL rPen-2 at 48 h was 13.2%, 62.4%, and 70.4%, respectively. DNA-specific fluorescent dye staining showed a high percentage of apoptosis on cancer cells. Flow cytometry revealed that the apoptosis rate of HK-2, ACHN and A498 cells was 15.2%, 55.2%, and 61.5% at 48 h respectively, suggesting that rPen-2 induced higher apoptosis rate in cancer cells than in HK-2 cells. Laser confocal scanning microscopy demonstrated that the plasma membrane was the key site where rPen-2 interacted with and destroyed tumor cells. Scanning electron microscopy showed the morphologic changes of the cell membranes of kidney cancer cells treated with rPen-2. These results suggest that rPen-2 is a novel potential therapeutic agent that may be useful in treating kidney cancers.
Animals
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Antimicrobial Cationic Peptides
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genetics
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pharmacology
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Antineoplastic Agents
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pharmacology
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Apoptosis
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drug effects
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Arthropod Proteins
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genetics
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pharmacology
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Drug Screening Assays, Antitumor
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Humans
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Kidney Neoplasms
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drug therapy
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metabolism
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pathology
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Penaeidae
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genetics
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Recombinant Proteins
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genetics
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pharmacology