1.Photothermal effects of metallic carbon nanotubes on human breast cancer cells.
Jin HOU ; Man MI ; Ming WEI ; Wen-hui YI
Acta Pharmaceutica Sinica 2015;50(8):980-985
SWNTs are a mixture of 1/3 metallic SWNTs (m-SWNTs) and 2/3 semiconducting SWNTs (s-SWNTs). It is desirable to separate the metallic SWNTs from the semi-conducting ones. In this study m-SWNTs was separated by using a poly[(m-phenylenevinylene)-alt-(p-phenylenevinylene)] (PmPV) derivative and used as photo-thermal media instead of SWNTs. The separation effects of m-SWNTs were evaluated by Raman spectra, molecular modeling and TEM images. The effects of m-SWNTs on MCF-7 cell proliferation and apoptosis were evaluated with MTT assay and flow cytometry, respectively. m-SWNTs were separated with high purity. A strong inhibition of MCF-7 cell growth was observed with the m-SWNTs under near-infrared (NIR) light irradiation. Our results will be helpful for the potential applications of m-SWNTs in clinical photothermal cancer therapy.
Apoptosis
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Breast Neoplasms
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pathology
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Flow Cytometry
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Humans
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Light
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MCF-7 Cells
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drug effects
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Models, Molecular
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Nanotubes, Carbon
2.Rapid Selection of Halophilic Streptomonospora Strains by PCR-SSCP
Man CAI ; Xiao-Yang ZHI ; Jin-Yuan WU ; Shu-Kun TANG ; Wen-Jun LI ;
Microbiology 2008;0(09):-
To improve the efficiency of halophilic actinobacteria screening and carry out the rapid selection of targeted strains, we tested 34 strains of Streptomonospora by PCR-single strand conformation polymorphism analysis (PCR-SSCP) based on genus-specific primers for the PCR identification. This approach employs PCR with two pairs of primers located in the 16S rRNA sequence flanking two variable region, then build clustering tree according as SSCP data. Synchronously, we sequenced all the 16S rRNA partial sequences for these strains to verify them. The results showed that the PCR-SSCP analysis was an efficient, easy-to-handle and economic method for rapid selection of halophilic actinobacteria resources.
3.Effect of doxycycline on the proliferation of bovine corneal myofibroblast in vitro
Man-li, LIU ; Wen-jin, ZOU ; Ming-han, HUANG ; Jing, ZENG ; Xin-yu, FU ; Song, WANG
Chinese Journal of Experimental Ophthalmology 2013;(5):428-433
Background Doxycycline is a broad spectrum antibiotic,and it is frequently used in the treatment of ocular surface diseases.Objective The purpose of the present study was to investigate the effect of doxycycline on the inhibition of cell proliferation in bovine corneal myofibroblasts in vitro and assess its contribution to ocular surface repairing mechanism.Methods Six fresh bovine corneas were collected.The corneal stromal layer was isolated by two-step method of 1.0 g/L and 2.0 g/L collegenase-1.Isolated cells were plated at mantaryay culture flask in 10% FBS of RPMI-1640.Vimentin and alpha-smooth muscle actin (α-SMA) organization were evaluated by immunocytochemistry,and the cells with influoresccence staining for vimentin and α-SMA were identified as the corneal myofibroblasts.Doxycycline at the concentrations of 10,20,40,60,80 mg/L was added to the medium,respectively,in different concentrations of doxycycline groups.Dexamethasone (120 mg/L)was used in the same way in the positive control group,and no drug was used in the negative control group.Cell proliferation was evaluated by MTT and the cell cycle was analyzed by BD FACScan flow cytometer assay 24 hours and 48 hours after addition of any drug.Results The cells grew well and showed the positive response for vimentin and α-SMA.MTT assay showed that the A570values of bovine corneal myofibroblasts were gradually declined with the increase of the concentration of doxycycline and lapse of active time,showing statistically significant difference (Fconcentration =1233.778,P<0.001 ; Ftime =227.564,P < 0.001).And the difference between the two factors was also statistically significant (Ftime*concentration =51.656,P<0.001).Flow cytometry cell cycle analysis showed that 24 hours after 10,20,40,60,80 mg/L doxycycline treated,the perentage of of corneal myofibroblast cell in G0-G1 phase was 82.85%,84.36%,85.18%,87.12 % and 89.31%,showing significant increase in comparison with 63.89% of the negative control group (all P<0.05),and that of 40 mg/L doxycycline group was near the positive control group.Forty-eight hours after 10,20,40,60,80 mg/L doxycycline treated,the perentage of of corneal myofibroblast cell in G0-G1 phase was 82.78%,86.15%,88.23%,89.57%,93.00%,with significant increase in comparison with 70.17% of the negative control group (all P < 0.01),and that of 40 mg/L doxycycline group was near the positive control group.Conclusions The growth of the bovine corneal myofibroblasts is inhibited by doxycycline in time-and dosedependent manner in the range from 10 mg/L to 80 mg/L,and 40 mg/L of doxycycline has an obviously inhibitory action as 120 mg/L dexamethasone.
4.Optimation of the Fermentation Conditions for Human ApolipoproteinA-I Expression
Wen-Feng MA ; Man-Sheng DING ; Mei-Jin GUO ; Ying-Ping ZHUANG ; Ju CHU ; Si-Liang ZHANG ;
Microbiology 1992;0(06):-
Optimization of the fermentation condition for human apolipoproteinA-I expression in recombinant Escherichia coli was investigated. The recombinant plasmid pBV220-ApoA-I was transformed respectively into different E.coli hosts such as JM109, BL21(DE3),DH5?, BMH7118,and TG1. The best host E.coli was DH5? in which the recombinant ApoA-I expression percentage was 21.2% corresponding to that in BL21(DE3) in flask shaker cultivation,while the ApoA-I expressed percentage in E.coli TG1 was 11%.Fed-batch cultivation was performed in FMG-5L fermentor,the optimum fermentation cultivation conditions were as following :optimum pH value was 7.0 in growth phase and 7.4 in the expression phase. The initial glucose concentration in batch phase was 3 g?L -1.The optimum C/N ratio was 2∶1.The recombinant ApoA-I reached about 40% of the total protein, and concentration of ApoA-I was 2.86 g?L -1.
5.Study on the Recombinant Human ApoA-I_Milano High Density and High Expression by Two Temperature-Shifted Induction in Escherichia coli
Wen-Feng MA ; Ying-Ping ZHUANG ; Mei-Jin GUO ; Man-Sheng DING ; Ju CHU ; Si-Liang ZHANG ;
Microbiology 1992;0(02):-
The temperature effect on the recombinant protein production formation was investigated in present study. The culture temperature of growth phase is 30℃, and the culture temperature of induction phase was arranged according to three modes. Hign cell-density and high expression culture of E.coli to product recombinant human apolipoprotein A-I Milano by two temperature-shifted induction . Two temperature-shifted induction was carried out high density and high expression recombinant human ApoA-1 Milano. The recombinant protein ApoA-I Milano reached 4.8 g?L -1 with the final cell density of OD 600 150. And the two temperature-shifted induction avoided the acetic acid successfully to the influence of the high density and high expression. Two temperature-shifted induction was viable in high density culture and high expression of heterogenous protein in recombination E.coli.The sduty provides a basic work for production of recombinant ApoA-I Milano in scale.
6.Comparative analysis of conventional pulmonary function test results in children with asthma or cough variant asthma.
Jie YUAN ; Shu-Hua AN ; Wen-Jie GAO ; Wen-Jin DU ; Jun-Feng SUN ; Man ZHANG ; Cong-Zhuo YAO
Chinese Journal of Contemporary Pediatrics 2013;15(3):171-174
OBJECTIVETo compare the conventional pulmonary function test results of children with asthma or cough variant asthma (CVA).
METHODSA total of 140 children, who were diagnosed with asthma or CVA from May 2010 to May 2011, were divided into acute asthma attack (n=50), asthma remission (n=50) and CVA groups (n=40); 30 healthy children were included as a control group. The forced vital capacity (FVC), forced expiratory volume in one second (FEV1), peak expiratory flow (PEF), forced expiratory flow after 25% of vital capacity has been expelled (FEF25), forced expiratory flow after 50% of vital capacity has been expelled (FEF50), forced expiratory flow after 75% of vital capacity has been expelled (FEF75) and maximal midexpiratory flow (MMEF75/25) were measured.
RESULTSThe mean percent predicted values of all the above indices were lower than 80% in the acute asthma attack group, with FEF50, FEF75 and MMEF75/25 declining markedly; the mean percent predicted values of FEF75 and MMEF75/25 were lower than 80% in the CVA group. All the pulmonary function indices in the acute asthma attack group were lower than those in the control group. The mean percent predicted values of FVC, FEV1, FEF25 and MMEF75/25 in the asthma remission and CVA groups were lower than in the control group. All the pulmonary function indices in the acute asthma attack group were lower than in the asthma remission and CVA groups, but there were no significant differences between the asthma remission and CVA groups.
CONCLUSIONSThere is small and large airway dysfunction, particularly small airway dysfunction, in children with acute asthma attack. Children with CVA present mainly with mild small airway dysfunction, as do those with asthma in remission.
Asthma ; physiopathology ; Child ; Cough ; physiopathology ; Female ; Forced Expiratory Volume ; Humans ; Lung ; physiopathology ; Male ; Vital Capacity
7.Study on clinical effect enhancement of acupuncture for depression with chronic pain treated with SSRI antidepressants.
Yi LIU ; Yong-Hua ZHANG ; Man JIN ; Wen-Juan LIU
Chinese Acupuncture & Moxibustion 2013;33(8):689-691
OBJECTIVETo compare the difference in the clinical efficacy on depression with chronic pain between the simple medication of selective serotonin uptake inhibition antidepressants (SSRIs) and the combined therapy of acupuncture and SSRIs.
METHODSNinety cases of depression were randomized into an acupuncture + medication group (combined therapy of acupuncture and western medication) and a medication group (simple medication), 45 cases in each one. In the medication group, the western medicine was adopted. According to the condition of disease, SSRIs was prescribed such as fluoxetine and paroxetine. In the acupuncture + medication group, on the basic treatment of western medication, acupuncture was applied to Baihui (GV 20), Yintang (GV 29), Shenting (BL 24), Fengchi (GB 20), etc. HAMD scale, VAS and SERS were used to evaluate the clinical efficacy in terms of depression symptoms, pain and side reactions of antidepressants before treatment and in the 1st, 2nd and 4th weeks after treatment in the patients and the differences between the two groups.
RESULTSThe differences in HAMD and VAS scores were not apparent before treatment between the two groups (P>0.05). HAMD score and VAS score in the 1st, 2nd and 4th weeks of treatment in the acupuncture + medication group were lower apparently than those in the medication group (all P<0.01). SERS score in the 1st, 2nd and 4th weeks of treatment in the acupuncture + medication group was lower apparently than that in the medication group (all P<0.01).
CONCLUSIONAcupuncture enhances the clinical efficacy on depression with chronic pain treated with SSRI antidepressants and reduces the adverse reactions.
Acupuncture Therapy ; Adolescent ; Adult ; Antidepressive Agents ; therapeutic use ; Combined Modality Therapy ; Depression ; drug therapy ; therapy ; Female ; Humans ; Male ; Middle Aged ; Serotonin Uptake Inhibitors ; therapeutic use ; Treatment Outcome ; Young Adult
8.Phentolamine antagonizes the effects of norepinephrine on the activity of pain-related neurons in the parafascicular nucleus of morphine-dependent rats.
Xiu-dong JIN ; Yan-zhong GUAN ; Shu-jie ZHANG ; Man-ying XU ; Wen-jie YUE
Journal of Southern Medical University 2008;28(2):266-268
OBJECTIVETo examine the antagonization of phentolamine against the effects of norepinephrine (NE) on the activity of pain-related neurons in the parafascicular nucleus of morphine-dependent rats.
METHODSElectric impulses were applied as nociceptive stimulus to the right sciatic nerve of morphine-dependent rats, and the discharges of the pain-related neurons in the parafascicular nucleus were recorded by extracellular recording method with glass microelectrodes.
RESULTSIntracerebroventricular injection of norepinephrine resulted in the inhibition of evoked response of the pain-excited neurons as well as the excitation of evoked response of the pain-inhibiting neurons. Both the inhibitory effect on the electric discharges of the pain-excited neurons and the excitatory effect on the pain-inhibiting neurons of norepinephrine were almost completely blocked by intracerebroventricular administration of phentolamine.
CONCLUSIONPhentolamine antagonizes the inhibitory effect of norepinephrine on the activity of pain-related neurons in the parafascicular nucleus in morphine-dependent rats, and norepinephrine may play an important role in the integration of the pain signal through the alpha-receptors.
Animals ; Drug Antagonism ; Electrophysiology ; Intralaminar Thalamic Nuclei ; cytology ; drug effects ; Neurons ; drug effects ; Norepinephrine ; antagonists & inhibitors ; pharmacology ; Pain ; physiopathology ; Phentolamine ; pharmacology ; Rats ; Rats, Wistar
9.Effects of hydrocortisone sodium succinate on sodium current in human and guinea pig cardiac myocytes.
Zhuang-li HU ; Hui LIU ; Yan HU ; De-yong ZHANG ; Zong-quan SUN ; Man-wen JIN
Acta Pharmaceutica Sinica 2004;39(4):250-253
AIMTo study the effects of hydrocortisone sodium succinate on sodium current in human atrial myocytes and in guinea pig ventricular myocytes.
METHODSSingle cardiac myocytes were isolated by enzyme. The effects of hydrocortisone sodium succinate on sodium current (INa) were assessed by applying whole-cell patch clamp techniques.
RESULTSHydrocortisone sodium succinate (1, 3, 10 micromol x L(-1)) was shown to inhibit INa of both human atrial myocytes and guinea pig ventricular myocytes in concentration dependent manner and the IC50 were 6.97 and 8.74 micromol x L(-1), respectively. The inhibition effects acted quickly (1-3 min) and the maximal activating voltage of INa was not changed in both human and guinea pig cardiac myocytes.
CONCLUSIONHydrocortisone sodium succinate can exhibit inhibitory effects on INa in both human and guinea pig cardiac myocytes, and its inhibitory effects act rapidly, which are not consistent with genomic effects, so there may be nongenomic effects.
Adolescent ; Adult ; Animals ; Cell Separation ; Child ; Child, Preschool ; Guinea Pigs ; Heart Atria ; pathology ; Heart Defects, Congenital ; pathology ; Heart Ventricles ; cytology ; Humans ; Hydrocortisone ; analogs & derivatives ; pharmacology ; Myocytes, Cardiac ; drug effects ; physiology ; Patch-Clamp Techniques ; Sodium Channels ; drug effects
10.Comparative metabonomics study on urine in rat treated by Angelica sinensis volatile oil.
Jin-Xia LI ; Man ZHANG ; Li-Bo SUN ; Ling ZHANG ; Wen-Quan ZHANG ; Hai-Fu ZHAO ; Peng-Ling LI ; Yong-Li HUA ; Peng JI ; Yan-Ming WEI
China Journal of Chinese Materia Medica 2014;39(7):1293-1299
Metabonomics was employed to investigate the effect of Angelica sinensis volatile oil (ASVO) to the endogenous metabolites of normal rats, and to reveal the possible ways of metabolism in rats caused by ASVO. The fifty male Waster rats were randomly divided into five groups (each consists of 10 rats), such as control group, high dose group of ASVO, middle dose group of ASVO, low dose group of ASVO, and Aspirin group. They were given 0.9% saline, 0.352 mL x kg(-1) ASVO, 0.176 mL x kg(-1) ASVO, 0.088 mL x kg(-1) ASVO and ASP respectively with the equal volume of 0.2 mL. Drugs and vehicle were given for 3 successive days. The urine was collected at 12, 24, 36, 48 h after modeling with metabolic cages. Rat urine metabolic fingerprint in different stages was analyzed using GC-MS, based on which the principal component analysis (PCA)and orthogonal partial least-squares discriminant analysis (OPLS-DA) models were established for metabonomic analysis. Potential biomarkers were screened by using variable importance in the projection (VIP) and T test. It was revealed that the middle dose of ASVO at 36 h induces a substantial change in rat urine. Compared with control group, seven kinds of endogenous metabolites in ASP group and ASVO group change significantly (P < 0.05), among which aconitic acid, succinic acid, citric acid, alpha-ketone glutaric acid, glycine and malic acid content had an upward trend (P < 0.05) and prostaglandin content had a downward trend (P < 0.01). The mechanism of ASVO and ASP have the similarity. It is likely that ASVO intervenes the metabolic process by affecting the energy, amino acid and lipid metabolism. Our work also indicates that rats administrated with ASVO can increase the energy metabolism of the body, induce the production of inflammatory substances and strengthen the body's immune ability. The result has also provide a proof for futher interpret ASVO pharmacological effects.
Angelica sinensis
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chemistry
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metabolism
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Animals
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Dose-Response Relationship, Drug
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Drugs, Chinese Herbal
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analysis
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metabolism
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pharmacology
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Energy Metabolism
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drug effects
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Lipid Metabolism
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drug effects
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Male
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Metabolomics
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Oils, Volatile
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analysis
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metabolism
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pharmacology
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Plant Oils
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analysis
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metabolism
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pharmacology
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Rats
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Rats, Wistar
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Urine
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chemistry