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1.
Chinese Journal of Otorhinolaryngology Head and Neck Surgery ; (12): 47-51, 2023.
Article in Chinese | WPRIM | ID: wpr-971406

ABSTRACT

Objective: To investigate the feasibility of only surgical resection for nasal vestibular squamous cell carcinoma and the efficacy of perforator flap of ipsilateral nasolabial sulcus in repairing postoperative defects. Methods: The clinical data of 8 cases with squamous cell carcinoma of the nasal vestibule who admitted to Department of Facial Plastic and Reconstructive Surgery, Eye & ENT Hospital, Fudan University were analyzed, including 6 males and 2 females, aged from 38 to 75 years. The tumor of the nasal vestibule was eradicated in time after making definite diagnosis of lesions, then the perforators flap of the ipsilateral nasolabial sulcus was used for repairment, without performing further chemotherapy or radiotherapy after surgery. The tumor recurrence, facial appearance, nostril form, donor area scar, nasal ventilation function, and cutaneous sensation were evaluated after surgery. Descriptive analysis was used in this research. Results: There were 2 cases of stage T1 and 6 cases of stage T2 in 8 cases. After 32 to 45 months of following-up, no recurrence accurred and all the flaps survived well. However, there was about 2 mm necrosis of the transplanted flap in the lateral foot of the alar in one case, which was healed well by carrying out wound care after 10 d. And the dark color flap was occurred in another case, showing the flap's backflow trouble, yet it was improved with addressing timely during 5 d postoperation. Pincusion-like deformity of the transplanted flap occurred in 4 cases (50%), which subsided gradually after 6 months. The morphology of the anterior nostril was altered in 4 cases (50%), but there was no ventilation trouble and no need for addressment in any case. The postoperative facial appearance was rated as excellentor good with hidden scar in the donor site, and the sensation of the transplanted flaps was indistinct from the surrounding tissue after 3 months. Conclusions: Surgical resection of nasal vestibular squamous cell carcinoma with tumor stage T1-2 is a feasible treatment. And it is the one of the best reconstructive methods of the perforator flap of the ipsilateral nasolabial sulcus to repair the deformities after the surgery.


Subject(s)
Male , Female , Humans , Perforator Flap/transplantation , Cicatrix/surgery , Neoplasm Recurrence, Local/surgery , Plastic Surgery Procedures , Carcinoma, Squamous Cell/surgery , Skin Transplantation/methods , Treatment Outcome
2.
Journal of Experimental Hematology ; (6): 747-752, 2019.
Article in Chinese | WPRIM | ID: wpr-771890

ABSTRACT

OBJECTIVE@#To establish a method for detecting the exosomal PML-RARA fusion gene expression by droplet digital PCR (ddPCR).@*METHODS@#By using Taqman probe-based ddPCR technique, the method that able to detect both long and short isoforms of PML-RARA fusion gene transcripts was established. RNA from PML-RARA negative cell line HL-60 as negative control was used to set the limit of blank (LOB), while the RNA from PML-RARA positive cell line NB4 and the recombinant plasmid pSG5-PML-RARA(S) were used to set the limit of detection (LOD) for long and short PML-RARA transcripts, respectively. Furtherly, the expression of exosomal PML-RARA fusion gene in NB4 cell culture supernatant and serum of patients with acute promyelocytic leukemia (APL) was analyzed by ddPCR technique.@*RESULTS@#The LOB of ddPCR assay for long and short PML-RARA transcripts were 0.0725 and 0.083 copies per microliter of PCR reaction system, respectively, while the LOD of long and short PML-RARA transcripts were 0.19 and 0.21 copies per microliter of PCR reaction system, respectively. In addition, the expression of exosomal PML-RARA fusion gene derived from both NB4 cell culture supernatant and serum of APL patients was successfully detected.@*CONCLUSION@#A ddPCR-based technique for detecting fusion gene transcripts has been established, which can be used to analyze absolute quantification in the minimal quantity of PML-RARA transcripts derived from exosomes. It suggests the possibility of this technique to non-invasively and dynamicly monitore the exosomal PML-RARA transcripts from APL patients' serum.


Subject(s)
Humans , Exosomes , Gene Expression , Leukemia, Promyelocytic, Acute , Oncogene Proteins, Fusion , Polymerase Chain Reaction , Protein Isoforms
3.
Chinese Journal of Preventive Medicine ; (12): 1023-1028, 2018.
Article in Chinese | WPRIM | ID: wpr-807566

ABSTRACT

Objective@#This study was designed to identify the relative factors, bacteriological profile and their antibiotic susceptibility pattern of neonatal sepsis.@*Methods@#A retrospective survey was conducted on the clinical information, pathogen identification and antibiotic sensitivity results of 425 newborns with neonatal sepsis admitted to Nanjing Maternity and Child Health Care Hospital from 2010 to 2017. Of the 425 positive blood-cultures, 148 (34.82%) were early-onset neonatal sepsis (EOS) and 277 (65.18%) were late-onset neonatal sepsis (LOS). Clinical information and pathogen identification were compared between EOS and LOS. Antibiotic sensitivity of gram negative organisms (G-) and gram positive organisms (G+) were also detected.@*Results@#The rates of premature delivery (78.70%, n=218), low birth weights (67.15%, n=186) and cesarean delivery (59.57%, n=165) were significantly increased in LOS (P<0.05) compared with those rates in EOS, which were 41.89% (n=62), 37.84% (n=56) and 46.62% (n=69). Parturients fever (18.24%, n=27) and meconium like amniotic fluid (25.68%, n=38) were significantly increased in EOS (P<0.05) compared with those rates in LOS, which were 7.94% (n=22) and 5.42% (n=15). Among the identified pathogen, the incidence of G- and G+ bacteria were 216 (50.83%) and 201 (47.29%) respectively, and the rest was Candida glabrata (1.88%, n=8). Escherichia coli 68 (16.00%) was the predominant isolate followed by Klebsiella pneumoniae (13.18%, n=56). The detection rate of Hemolytic staphylococcus (10.81%, n=16) was significantly increased in EOS (P<0.001) compared with LOS (1.44%, n=4). However, the incidence of Klebsiella pneumonia (5.88%, n=44) was higher in LOS (P=0.024) compared with EOS (8.11%, n=12). Most of the gram positive isolates exhibited high resistance to penicillin (90.32%-100.00%) and cephalosporin group antibiotics (25.00%-100.00%). Similarly, the majority of the gram negative isolates showed higher resistance to ampicillin (83.33%-100.00%), but susceptible to aminoglycosides (0-11.76%), quinolones (0-17.65%) and β-lactams (0-5.88%).@*Conclusion@#Among the study population, the percent of preterm, low birth weight and cesarean section were higher in LOS while parturients fever and meconium-like amniotic fluid were higher in EOS. The pathogens with the highest detection rate were Escherichia coli and Klebsiella pneumoniae. The results of antibiotic susceptibility test showed that common pathogens had high resistance to commonly used antibacterial drugs.

4.
Chinese Journal of Analytical Chemistry ; (12): 217-224, 2018.
Article in Chinese | WPRIM | ID: wpr-692238

ABSTRACT

Bismuth modified boron doped diamond (BDD) film electrode was employed for simultaneous determination of trace ZnⅡ,CdⅡand PbⅡby anodic stripping voltammetry.BiⅢwas simultaneously in-situ deposited on bismuth modified boron doped diamond electrode with ZnⅡ,CdⅡ and PbⅡ by pre-concentration.In the presence of BiⅢ,the sensitivity for determination of ZnⅡ,CdⅡ and PbⅡ was remarkably enhanced.Influence factors such as bismuth concentration,boron doped concentrations of BDD electrode,pH,preconcentration potential were investigated and optimized.Under the optimal conditions,the stripping peak currents increased linearly with the increasing concentration of ZnⅡ,CdⅡ and PbⅡ in the range of 10-300 μg/L.The limit of detection was 0.56 μg/L for ZnⅡ,0.32 μg/L for CdⅡand 0.75 μg/L for PbⅡ (S/N=3),respectively.The interference experiments showed that common ions had little influence on the determination except CuⅡ.In addition,the developed electrode displayed a good repeatability.The method was successfully applied to determination of ZnⅡ,CdⅡ and PbⅡ in real water samples with the standard addition recoveries of 92.0%-114.0%.

5.
Chinese Journal of Tissue Engineering Research ; (53): 40-45, 2018.
Article in Chinese | WPRIM | ID: wpr-698337

ABSTRACT

BACKGROUND: Retrorsine (RS) is a chemical agent for the long-term inhibition of mature liver cell division and proliferation. OBJECTIVE: To establish a rat model of liver injury by combined use of RS and 1/3 partial hepatectomy, to observe the proliferation of liver cells and oval cells in rats after liver injury, and to discuss the relationship between liver regeneration and mature liver cells and oval cells after liver injury. METHODS: Thirty male Sprague-Dawely rats were randomized into two groups (n=15 per group): RS group and control group. Rats in the RS group were subjected to intraperitoneal injection of RS, 30 mg/kg, twice in total, with 2 weeks in between; and rats in the control group were injected physiological saline instead of RS. Four weeks after the last injection, the 1/3 partial hepatectomy was performed in two groups of rats. Liver pathological and morphological changes as well as cell proliferation were observed, and CK19 and C-kit immunohistochemical detections of the rat liver in the two groups were conducted at different time points after operation. RESULTS AND CONCLUSION: At 20 days after operation, the body mass of the RS group rats was still lower than the baseline, and the liver increase was obviously less than that in the control group; there was cell body swelling shown by hematoxylin-eosin staining, loose cytoplasm, extensive vacuoles degeneration of liver cells, and clustered or scattered oval cells around the portal area of small bile duct and in the hepatic lobule. However, in the control group, the body mass was close to the baseline, liver damage was lighter than that in the RS group, a large number of mature liver cells proliferated under BrdU Immunofluorescence at 20 days after operation; liver oval cells proliferated and distributed in the liver cell line at 14 days after operation, with morphology and immunohistochemical markers consistent with oval cells in the RS group. These findings indicate that the rat model of acute liver injury is successfully established by combining retrorsine with 1/3 partial hepatectomy, liver poisoning and the proliferation of liver stem cells and mature liver cells after poisoning can be seen in the experiment, which firmly confirm that liver cell renewal and regeneration after injury is accredited to the combined action of liver stem cells in liver basin and mature liver cells.

6.
Journal of Experimental Hematology ; (6): 346-352, 2017.
Article in Chinese | WPRIM | ID: wpr-311540

ABSTRACT

<p><b>OBJECTIVE</b>To explore the growth inhibitory effect of quercetin on imatinib-resistant chronic myeloid leukemia cell lines and to clarify its involved mechanisms.</p><p><b>METHODS</b>The cell viability was detected by trypan blue Staining, percentage of apoptotic cells and cell cycle distribution were detected by flow cytometry, the protein expression was detected by Western blot.</p><p><b>RESULTS</b>Both inhibitory effect of proliferation and apoptosis-inducing effect were similar between the imatinib-resistant and -sensitive cell lines treated with 25 µmol/L quercetin for 24 hours and with arrest of cell cycle at G/M phase. Quercetin could not change the expression of BCR-ABL. The expression of γ-H2AX was markedly enhanced and the phosphorylation of JNK up-regulated by quercetin in both imatinib-resistant and imatinib-sensitive cell lines.</p><p><b>CONCLUSION</b>The growth of imatinib-resistant cells can be inhibited by quercetin, and the apoptosis of cells can be induced by quercetin, which may be related to cell cycle arrest in G/M. The DNA damage and up-regulation of p-JNK may be involved in these processes.</p>

7.
Journal of Medical Biomechanics ; (6): E369-E375, 2016.
Article in Chinese | WPRIM | ID: wpr-804042

ABSTRACT

An abdominal aortic aneurysm (AAA) is a vascular pathology associated with localized and balloon-like dilatations of abdominal aorta. An untreated AAA may lead to an eventual rupture with a high mortality rate. In recent studies, the biomechanics of AAA has been widely used to assess the rupture risk in clinic. In this review paper, biomechanical testing methods on intraluminal thrombi and AAA are discussed, so as to fully understand biomechanical properties of intraluminal thrombi and aneurysmal tissues, as well as the influence of mechanical property changes on the AAA growth and remodeling under pathological environment. Then representative research findings on prediction of rupture risk by a series of experimental and computational biomechanical methods are reviewed, including finite element analysis on stress distributions on AAA wall, assessment of rupture risk index and judgment of rupture locations. The relevant microstructural changes caused by thrombus aging are described in detail, and the current situation of biomechanical studies on AAA and future challenges are briefly summarized.

8.
Journal of Experimental Hematology ; (6): 966-970, 2015.
Article in Chinese | WPRIM | ID: wpr-357237

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the biological effects of sertraline, one of psychotropic drugs, on actue myeloid leukemia cell line Kasumi-1.</p><p><b>METHODS</b>Cells were treated by different concentrations of sertraline for different times. The effects of sertraline were evaluated by cell growth, cell morphology, cell cycle distribution and markers of cell apoptosis, respectively. Western blot was used to detect the expression change of related proteins.</p><p><b>RESULTS</b>Sertraline could inhibit cell proliferation and induce apoptosis. After treatment with 15 µmol/L and 20 µmol/L sertraline for 24 h, the inhibitory rate of Kasumi-1 cell proliferation was (19.00 ± 7.37)% and (47.90 ± 11.19)%, respectively. Meanwhile, compared with the control group, the percentage of Annexin V positive cells in Kasumi-1 cells treated with sertraline for 24 h raised obviously from (9.71 ± 2.12)% to (20.54 ± 2.52)% and (45.37 ± 7.88)% (P < 0.01), respectively. The cells were arrested in G0/G1 and G2/M phase. In addition, it was found that sertraline could also down-regulate the level of translationally controlled tumor protein (TCTP) in Kasumi-1 cells.</p><p><b>CONCLUSION</b>Sertraline can significantly induce the apoptosis of Kasumi-1 cells, that probably is associated with the down-regulation of TCTP protein expression.</p>


Subject(s)
Humans , Apoptosis , Cell Cycle , Cell Line, Tumor , Cell Proliferation , Down-Regulation , Sertraline
9.
National Journal of Andrology ; (12): 325-329, 2015.
Article in Chinese | WPRIM | ID: wpr-319500

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect and safety of extracorporeal shock wave (ESW) in the treatment of pain symptom of type III B prostatitis.</p><p><b>METHODS</b>We treated 50 cases of type III B prostatitis by ESW once a week for 4 weeks. Then we evaluated the clinical effect and safety of the therapy based on the NIH-CPSI scores, visual analogue scale (VAS) scores, IIEF-5 scores, prostate volume and morphous, state of urination, color of urine, results of routine semen analysis, and changes of cytokines (IL-6, TNF-α and IL-1β) in expressed prostatic secretion (EPS).</p><p><b>RESULTS</b>All the patients successfully accomplished the treatment. Compared with the baseline, decreases were observed after 4 weeks of cytokine treatment in the pain scores (14. 61 ± 1. 82 vs 9. 36 ± 1. 47, P <0. 01), urination symptom scores (4. 59 ± 1. 01 vs.4. 66 ± 0. 89, P >0. 05) , quality of life scores (6. 51 ± 1. 03 vs 4. 56 ± 1. 02, P <0. 01), NIH-CPSI (25. 43 ± 1. 72 vs 18. 28 ± 2. 32, P <0. 01 ), and VAS (6. 59 ± 1. 10 vs 3. 02 ± 1. 07, P < 0. 01). The concentration of IL-6 in the EPS was significantly increased ([55.82 ± 6. 28] vs [86.59 ± 4. 55] ng/ml, P <0. 01) , while the level of TNF-α ([3.89 ± 0. 12] vs [3. 19 ± 0.22] ng/ml, P<0.01) and that of IL-1β ([3.21 ± 1.01] vs [1.48 ± 0.95] ng/ml, P< 0. 01) remarkably reduced after treatment. However, there were no statistically significant differences in IIEF-5 scores (18. 58 ± 2. 03 vs 18. 51 1. 89, P >0. 05) or various sperm parameters before and after treatment (P >0. 05). And no significant changes were observed in the prostate volume, morphous or internal echoes.</p><p><b>CONCLUSION</b>The ESW therapy is effective and safe for the pain symptom of type III B prostatitis.</p>


Subject(s)
Adult , Humans , Male , Middle Aged , Body Fluids , Interleukin-1beta , Metabolism , Interleukin-6 , Metabolism , Pain , Metabolism , Pain Management , Methods , Prostatitis , Metabolism , Therapeutics , Quality of Life , Spermatozoa , Physiology , Tumor Necrosis Factor-alpha , Metabolism , Ultrasonic Therapy , Methods , Urine
10.
Journal of Medical Postgraduates ; (12): 688-691, 2015.
Article in Chinese | WPRIM | ID: wpr-462445

ABSTRACT

Objective Few researches are reported on the association of integrinαvβ6 with vaginal mucous infection and de-fense.This study aimed to investigate the effects of IL-6, TGF-β, and IFN-γon the expression of integrinαvβ6 in the vaginal epithelial cell line VK2/E6E7. Methods Immortalized human vaginal epi-thelial cells (VK2/E6E7) were cultured in vitro and treated with gra-dient concentrations of IL-6 (1, 10, 50, and 100 ng/mL), TGF-β(0.1, 1, 10, and 100 ng/mL), and IFN-γ(50, 500, 2500, and 5000 U/L) , respectively.After 48 hours, the cells were collected and total RNA extracted by the Trizol method to be reversely tran-scribed to cDNA. The expressions of integrin αand β6 subunit mRNA were detected by real-time quantitative PCR. Results In the IL-6-treated VK2/E6E7 cells, the integrin αand β6 subunit mRNA expressions were significantly lower in the 1 ng/mL and 10 ng/mL groups but remarkably higher in the 100 ng/mL group (1.14 ±0.12 and 1.37 ±0.25) than in the blank control (1.00 ±0.09) (P<0.05), and only the expression ofβ6 subunit mRNA was elevated in the 50 ng/mL group (P<0.05), with the expressions increased in a concentration-dependent manner.In the TGF-βtreated VK2/E6E7 cells, the expressions of integrinαandβ6 subunit mRNA were significantly lower in the 0.1 ng/mL and 1 ng/mL groups but remarkably higher in the 100 ng/mL group than in the blank control (1.00 ±0.09) (P<0.01), and only the expression ofβ6 subunit mRNA was elevated in the 10 ng/mL group (4.31 ±0.78, P<0.01), with the expressions increased in a concentration-dependent manner.In the IFN-γtreated VK2/E6E7 cells, the expressions of both integrinαandβ6 subunit mRNA were significantly lower in the 50 U/L, 500 U/L, 2500 U/L, and 5000 U/L groups than in the blank control (all P<0.01). Conclusion IL-6 and TGF-βhave an inhibitory effect the expression ofαvβ6 in VK2/E6E7 cells at low concentrations, which gradually diminishes with the increased concentration of inflammatory factors.In the early stage of inflammation, IFN-γcan effectively suppress the expression ofαvβ6 at a high concentration.However, with the progression of inflammation and decrease of its concentration, IFN-γloses its inhibi-tory effect and therefore does not help inflammation control.

11.
Chinese Traditional and Herbal Drugs ; (24): 3091-3095, 2015.
Article in Chinese | WPRIM | ID: wpr-853929

ABSTRACT

Objective: To compare the contents of three homoisoflavones and their anti-oxidative activity of Ophiopogon japonicus in Sichuan (OJS) and Zhejiang (OJZ). Methods: The determination was performed on ACQUITYTM UPLC BEH C18 column (100 mm×2.1 mm, 1.7 μm), the Ultra Performance Liquid Chromatography-Photo-Diode Array (UPLC-PDA) technology was applied, and the mobile phase consisted of acetonitrile and 0.2% formic acid aqueous solution (55:45) with gradient elution program, flow rate was 0.20 mL/min with 2 μL of sample quantity at 296 nm, and the anti-DPPH radical efficiency of water extract from Ophiopogonis Radix were evaluated by UV-photometer. Results: The average values of methylophiopogonone A, methylophiopogonanone A, and methylophiopogonanone B were (3.06±0.54), (40.10±5.63), and (29.51±5.06) μg/g in OJS, respectively; while those in OJZ were (9.22±3.52), (106.63±27.56), and (256.97±61.79) μg/g, separately. The IC50 value was 16.59 mg/mL in OJS, while that in OJZ was 14.48 mg/mL. The IC50 value of positive control VC was 7.06 μg/mL. Conclution: Compared with OJS, the contents of homoisoflavones in OJZ are higher, and the anti-radical efficiency of water extract from OJZ is stronger. It provides the basis for the quality evaluation and geo-authentic research of Ophiopogonis Radix.

12.
Acta Physiologica Sinica ; (6): 151-157, 2014.
Article in Chinese | WPRIM | ID: wpr-297506

ABSTRACT

The aim of the present study was to investigate the effect of precursor brain-derived neurotrophic factor (proBDNF) on survival and neurite outgrowth of cultured rat spiral ganglion neurons (SGNs). Spiral ganglions (SG) were collected from postnatal day 5 Sprague Dawley (SD) rats, then enzymatically digested and suspended. Dissociated SGNs were plated on poly-D-lysine/laminin coated eight-well chamber plates and maintained at 37 °C for 4 h to promote the attachment of the neurons. Cultured SGNs were randomly divided into five groups: control group, BDNF group (BDNF 10 ng/mL), C10 group (proBDNF 10 ng/mL), C50 group (proBDNF 50 ng/mL), and C100 group (proBDNF 100 ng/mL). All groups were incubated in a serum-free medium. 48 h after incubation, SGNs were fixed and stained for βIII tubulin. Immunostaining of the cultured SGNs showed that, compared with the control group, the cellular survival of C50 group and C100 group were significantly reduced (P < 0.001). Furthermore, surviving numbers of the three proBDNF-treated groups were all lower than the BDNF group. In order to assess the effect of proBDNF on cell morphology, SGNs were divided into two categories: SGNs with or without neurites. The results demonstrated that proBDNF significantly increased the proportions of SGNs without neurites in C10, C50 and C100 groups compared with that in control group (P < 0.001). In addition, c-Jun N-terminal kinase (JNK) inhibitor, SP600125 (20 μmol/L) significantly increased the surviving number of SGNs in C50 group. These results suggest that proBDNF reduces the survival rate of cultured SGNs and inhibits the sprouting of neurites. Furthermore, the inhibition of JNK signaling attenuates the effect of proBDNF on SGNs survival.


Subject(s)
Animals , Rats , Axons , Physiology , Brain-Derived Neurotrophic Factor , Pharmacology , Cell Survival , Cells, Cultured , JNK Mitogen-Activated Protein Kinases , MAP Kinase Signaling System , Neurites , Physiology , Neurons , Cell Biology , Protein Precursors , Pharmacology , Rats, Sprague-Dawley , Spiral Ganglion , Cell Biology
13.
Acta Pharmaceutica Sinica ; (12): 1451-1456, 2014.
Article in Chinese | WPRIM | ID: wpr-299112

ABSTRACT

To evaluate the differences of Ophiopogonjaponicus from different cultivations, the metabolomics based method was conducted to compare the effects of Hangmaidong and Chuanmaidong (Chinese name) on plasma endogenous metabolites of normal rats. Data were collected by ultra performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-Q-TOF/MS), and were analyzed by multivariate statistical method, such as Principal Component Analysis and Orthogonal Signal Correction Partial Least Square Discriminant Analysis. Results revealed that the plasma metabolites profiling of low and middle dose group of Chuanmaidong were similar to the control group, but different from the high dose group obviously. Meanwhile the high, middle and low dose groups of Hangmaidong were different from control group notably, and the difference is dose dependent. Lysophosphatidylcholines, the possible endogenous metabolites which contribute to the classification most significantly, are closely related to cardiovascular system diseases. Compared with the group of Chuanmaidong, Hangmaidong has greater impact on the plasma metabolic profiling of normal rats. Hangmaidong and Chuanmaidong showed significant differences pharmacodynamically.


Subject(s)
Animals , Rats , Biomarkers , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Pharmacology , Least-Squares Analysis , Mass Spectrometry , Metabolomics , Principal Component Analysis
14.
Journal of International Pharmaceutical Research ; (6): 200-205, 2014.
Article in Chinese | WPRIM | ID: wpr-845780

ABSTRACT

Objective To stably express rat mu-opioid receptor (rMOR) in PC12 cells with characteristics of neurons. Methods After the lentiviral vector pBPLV-rMOR-eGFP was constructed, the lentivirus was packaged and used to infect the PC12 cells. PC12 cells stably expressing rMOR was screened by the flow cytometry and the limiting dilution assay. The affinity and quantity of rMOR protein expressed in the PC12 were verified by radio-ligand binding assay. The function of rMOR was analyzed by cAMP overshooting. Results The eGFP protein in PC12-rMOR cells infected by the lentivirus could be clearly shown by the fluorescence microscope. Cell lines grew normally after every clone was enlargedly cultured. The affinity (Kd) and quantity (Bmax) values of rMOR were (0.51 ± 0.07) nmol/L and (1.58 ± 0.15)pmol/mg protein respectively in 3H-diprenorphine binding assay. The cAMP content increased (255 ±25.2) % after naloxone precipitated in chronic morphine-treated cells. Exogenous agmatine could dose-dependently inhibit the overshooting of cAMP by naloxone precipitated. Conclusion We have successfully established the PC12 cell model co-expressing stably rMOR and I1 style imidazoline receptor(I1R) without α2 adrenergic receptor, expressing properties of neurons, which is a good cell model in vitro for investigating the neural molecular mechanism of opioid addition and regulating the opioid receptor function by the system of agmatine and I 1 Rin the future.

15.
Journal of Experimental Hematology ; (6): 1356-1360, 2012.
Article in Chinese | WPRIM | ID: wpr-325260

ABSTRACT

This study was aimed to further explore the apoptosis-inducing effect of bortezomib combined with cytarabine (Ara-C) on U937 cell line. Proliferation and apoptosis in U937 cells treated with bortezomib and/or Ara-C were assessed by cell count. Cell cycle distribution and reactive oxygen species (ROS) production level were measured by using flow cytometry. Cell signaling pathway related to apoptosis was analyzed by Western blot. The results showed that 10 nmol/L bortezomib combined with 50 nmol/L Ara-C significantly inhibited U937 cell proliferation. These two drug combination synergistically induced apoptosis in U937 cells, significantly increased cellular ROS level, and up-regulated the expression of phosphorylated form of JNK and P38 and down-regulated phosphorylation of ERK. It is concluded that the apoptosis of U937 cells synergistically induced by bortezomib combined with low concentration Ara-C is possibly associated with up-regulation of phosphorylated form of JNK, P38 and down-regulation of phosphorylation of ERK induced by increase of ROS, resulting in decrease of mitochondrial potential.


Subject(s)
Humans , Apoptosis , Boronic Acids , Pharmacology , Bortezomib , Cytarabine , Pharmacology , Drug Synergism , Gene Expression Regulation, Neoplastic , MAP Kinase Signaling System , Phosphorylation , Pyrazines , Pharmacology , Reactive Oxygen Species , Metabolism , U937 Cells
16.
Journal of Experimental Hematology ; (6): 554-557, 2012.
Article in Chinese | WPRIM | ID: wpr-263351

ABSTRACT

This study was aimed to explore the effect of bortezomib and low concentration cytarabine (Ara-C) on proliferation and apoptosis in U937 cell line and its mechanism. The proliferation and apoptosis of U937 cells treated with bortezomib (10 nmol/L) and(or) Ara-C (50 nmol/L) were observed by cell count, cell morphology, flow cytometry and Western blot. The results showed that bortezomib and Ara-C alone inhibited U937 cell proliferation. The inhibitory effect was enhanced by combination of these two drugs, the inhibitory rates of U937 cell proliferation were (55.00 ± 2.81)% and (70.02 ± 3.33)% after treatment for 24 h and 48 h, respectively. Bortezomib and Ara-C synergistically induced apoptosis and decreased mitochondrial membrane potential in U937 cells. The percentage of Rhodamin123 positive cells was (38.70 ± 1.54)%. Bortezomib and Ara-C also synergistically induced activation of caspase-9, caspase-8 and caspase-3. It is concluded that the bortezomib and low concentration Ara-C synergistically induced apoptosis in U937 cells, mainly through mitochondrial pathway, and possibly through death receptor pathway.


Subject(s)
Humans , Apoptosis , Boronic Acids , Pharmacology , Bortezomib , Cell Cycle , Cytarabine , Pharmacology , Pyrazines , Pharmacology , U937 Cells
17.
Chinese Journal of Industrial Hygiene and Occupational Diseases ; (12): 87-93, 2011.
Article in Chinese | WPRIM | ID: wpr-272651

ABSTRACT

<p><b>OBJECTIVE</b>To study the curative effects of pirfenidone (PF) on pulmonary fibrosis induced by paraquat (PQ) in mice and to provide the theoretical basis for clinical treatment.</p><p><b>METHODS</b>Ninety adult healthy male ICR mice were randomly divided into six groups: control group, PQ group, 2 mg/kg Dexamethasone group, 25 mg/kg PF group, 50 mg/kg PF group and 100 mg/kg PF group, there were 15 mice in each group. The corresponding volume of normal saline was given to the each mouse in control group according to the weight, after 2 h 0.1% CMC was given to the each mouse of control group one time by intragastric administration, then the CMC was administrated at regular time until sacrifice. All mice for other 5 groups were exposed to 100 mg/kg PQ by intragastric administration. At 2 h after exposure to PQ, 0.02 ml/10 g dexamethasone and 25, 50, 100 mg/kg PF were given to mice for dexamethasone group and for 3 PF groups by intragastric administration each day for 49 days, respectively. The lung coefficient was calculated and pathological changes of lung tissue were observed by HE staining for each mouse. The hydroxyproline (HYP) level in lung tissue was measured for each mouse. The mRNA level of and the protein level of TGF-β(1) in lung tissue for each mouse were determined, and the protein level of TGF-β(1) in the bronchus-alveolus lavage fluid (BALF) of each mouse was detected.</p><p><b>RESULTS</b>The survival rates on the 3rd day in PQ group, 3 PF groups and dexamethasone group were 53.33%, 46.67%, 73.33%, 86.67% and 80%, respectively. The survival rates on the 3rd day in dexamethasone group, 50 mg/kg and 100 mg/kg PF groups were significantly higher than those of PQ group and 25 mg/kg PF group (P < 0.05). The lung coefficients of 3 PF groups were significantly lower than that of the PQ group (P < 0.05). The lung tissue HYP levels of dexamethasone group and 3 PF groups were 50.95 ± 11.65, 44.52 ± 9.48, 43.27 ± 6.01 and 40.82 ± 5.90 mg/g respectively, which were significantly lower than that (74.27 ± 3.68) of PQ group (P < 0.01). The TGF-β(1) protein levels of BALF in dexamethasone group, 50 and 100 mg/kg PF groups were 22.03 ± 7.27, 27.75 ± 5.84 and 21.31 ± 6.82 ng/ml respectively, which were significantly lower than that (52.52 ± 15.51) ng/ml of PQ group (P < 0.01) The expression level of TGF-β(1) mRNA in 100 mg/kg PF group decreased significantly, as compared with PQ group (P < 0.01).</p><p><b>CONCLUSION</b>PF could reduce the collagen deposition and pulmonary fibrosis induced by PQ in mice lungs.</p>


Subject(s)
Animals , Male , Mice , Disease Models, Animal , Lung , Metabolism , Pathology , Mice, Inbred ICR , Paraquat , Poisoning , Pulmonary Fibrosis , Drug Therapy , Pathology , Pyridones , Therapeutic Uses , Transforming Growth Factor beta , Metabolism
18.
Journal of Experimental Hematology ; (6): 31-35, 2010.
Article in Chinese | WPRIM | ID: wpr-328578

ABSTRACT

To investigate the molecular mechanisms of all-trans retinoic acid (ATRA)-induced rig-g gene expression and to better understand the signal transduction of ATRA during acute promyelocytic leukemia (APL) cell differentiation, the luciferase reporter assay, co-immunoprecipitation and chromatin immunoprecipitation were used to clarify the basic transcriptional factors, which directly initiated the expression of rig-g gene. The results showed that the expression of STAT2, IRF-9 and IRF-1 could be upregulated by ATRA with different kinetics in NB4 cells. IRF-9 was able to interact with STAT2 to form a complex, which could bind the rig-g gene promoter and trigger the rig-g expression. IRF-1 alone could also activate the reporter gene containing rig-g gene promoter, but C/EBPalpha could strongly inhibit this transcription activity of IRF-1. It is concluded that during ATRA-induced APL cell differentiation, IRF-1 is first upregulated by ATRA, and then IRF-1 increases the protein levels of IRF-9 and STAT2 with the downregulation of C/EBPalpha. The complex of IRF-9 and STAT2 is the primary transcriptional factor for rig-g gene induction. This study will be helpful for better understanding the signal transduction networks of ATRA during the course of APL cell differentiation.


Subject(s)
Humans , CCAAT-Enhancer-Binding Protein-alpha , Metabolism , Gene Expression Regulation, Leukemic , Genes, Regulator , Interferon Regulatory Factor-1 , Metabolism , Interferon-Stimulated Gene Factor 3, gamma Subunit , Metabolism , Intracellular Signaling Peptides and Proteins , Genetics , Leukemia, Promyelocytic, Acute , Genetics , STAT2 Transcription Factor , Metabolism , Signal Transduction , Tretinoin , Pharmacology , Tumor Cells, Cultured
19.
Chinese Journal of Medical Genetics ; (6): 255-258, 2010.
Article in Chinese | WPRIM | ID: wpr-349002

ABSTRACT

<p><b>OBJECTIVE</b>To study the regulatory role of interferon-stimulated response elements (ISREs) located on the retinoic acid-induced gene G (RIG-G) promoter in RIG-G expression.</p><p><b>METHODS</b>By using point mutation technique, the authors constructed the wide type and site mutant reporter gene plasmids according to the ISRE sequence on RIG-G promoter, and detected the functional activities by luciferase reporter assay.</p><p><b>RESULTS</b>Mutation in ISRE II alone had no obvious effect on the expression of the reporter gene, whereas mutation in ISRE I dramatically inhibited the transactivity of RIG-G promoter. Mutation in both ISRE I and ISRE II resulted in complete loss of its response to the transcription factors for the reporter gene.</p><p><b>CONCLUSION</b>Both ISRE I and ISRE II on the RIG-G promoter are the binding sites for the complex of transcription factors. They are required for RIG-G expression, and ISRE I has a preferential role over ISRE II.</p>


Subject(s)
Humans , Cell Line, Tumor , Interferon Regulatory Factor-1 , Genetics , Metabolism , Interferons , Genetics , Metabolism , Intracellular Signaling Peptides and Proteins , Genetics , Mutation , Promoter Regions, Genetic , Genetics , Response Elements , Genetics , STAT2 Transcription Factor , Genetics , Metabolism
20.
Journal of Experimental Hematology ; (6): 1159-1162, 2010.
Article in Chinese | WPRIM | ID: wpr-237573

ABSTRACT

This study was purposed to investigate the expression of ifi56 gene in the ATRA-induced acute promyelocytic leukemia (APL) NB4 cell differentiation and to construct the eukaryotic expression plasmid of ifi56 gene. RT-PCR was used to detect the expression of ifi56 in NB4 cells treated with ATRA for different time. Human ifi56 cDNA was amplified by RT-PCR and cloned into pEGFP-C1 vector, then was transfected into 293T cells. The expression of the recombinant protein in 293T cells was detected by Western blot. The localization of IFI56 protein was observed by fluorescence microscopy. The results showed that the ifi56 mRNA was almost undetectable in untreated NB4 cells, but it significantly increased after ATRA treatment for 72 hours. The cDNA fragment of ifi56 was inserted into the expressing plasmid pEGFP-C1 successfully. The expression of EGFP-IFI56 fusion protein with a molecular weight about 83 kD was detected by Western blot. The EGFP-IFI56 protein was localized in cytoplasm mainly. It is concluded that the expression of ifi56 is enhanced significantly when the differentiation of APL cells was induced by ATRA. Gene ifi56 is successfully cloned into eukaryotic expression vector and the fusion protein is expressed in the cytoplasm mainly.


Subject(s)
Humans , Carrier Proteins , Genetics , Cell Differentiation , Genetics , Cell Line, Tumor , Gene Expression , Genetic Vectors , Leukemia, Promyelocytic, Acute , Genetics , Recombinant Fusion Proteins , Genetics , Tretinoin , Pharmacology
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