2.Expression of Smad 7,Smurf 1 and Smurf 2 in basal cen carcinoma and squamous cell carcinoma
Ying LI ; Wei HE ; Yun-Zhi HE ; Hai HUANG ; Zi-Hua LIN ; Jun WU
Chinese Journal of Dermatology 2003;0(12):-
Objective To investigate the expression and significance of Smad 7,Smurf 1 and Smurf 2 in basal cell carcinoma and squamous cell carcinoma.Methods Biopsy specimens were resected from 14 patients with basal cell carcinoma,19 patients with squamous cell carcinoma and 30 normal controls.Quanti- tative real-time PCR and immunohistochemical techniques were utilized to assess the expression of Smad 7, Smurf 1 and Smurf 2 in these specimens.Results The gray scale for staining of Smad 7,Smurf 1 and Smurf 2 was 166.61?7.11,166.08?8.71,and 166.25?8.15 respectively in basal cell carcinoma,161.66?5.52,166.84?9.27,and 169.98?9.48 respectively in squamous cell carcinoma.The expression levels of Smad 7,Smurf 1 and Smurf 2 were all significantly increased in basal cell carcinoma and squamous cell car- cinoma in comparison with normal controls.Conclusions The over-expression of Smad 7,Smurf 1 and Smurf 2 may interfere with transforming growth factor?signaling transduction pathway through several links,therefore prevent the inhibitory effect of transforming growth factor?on epidermal proliferation,and accelerate the abnormal proliferation in above epidermal tumors.
3.Identification of differentially expressed microRNAs in the ovary of polycystic ovary syndrome with hyperandrogenism and insulin resistance.
Lin LIN ; Tao DU ; Jia HUANG ; Li-Li HUANG ; Dong-Zi YANG
Chinese Medical Journal 2015;128(2):169-174
BACKGROUNDPolycystic ovary syndrome (PCOS) is the commonest endocrinopathy in women of reproductive age. The patients often develop insulin resistance (IR) or hyperinsulinemia despite manifesting anovulation and signs of hyperandrogenism. The cause and effect relationship of hyperinsulinemia and hyperandrogenemia (HA) is still debated. Micro-ribonucleic acids (miRNAs) have recently been shown to play a role in regulation of ovarian function. Our current study focused on the altered expression of miRNAs with PCOS.
METHODSOvarian theca interna tissues were obtained from 10 PCOS patients and 8 controls that were non-PCOS and had normal insulin sensitivity undergoing laparoscopy and/or ovarian wedge resection. Total RNA of all samples was extracted. We studied the repertoire of miRNAs in both PCOS and non-PCOS women by microarray hybridization. Bioinformatic analysis was performed for predicting targets of the differentially expressed miRNAs. Furthermore, selected miRNAs were validated by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR).
RESULTSA total of 27 miRNAs were differentially expressed in PCOS patients with respect to the controls in our discovery evaluationand two (miR-92a and miR-92b) of them were significantly downregulated in PCOS women in followed validation (P < 0.05). Targets prediction revealed that miR-92a targeted both GATA family of zinc finger transcription factor GATA-binding factor 6 (GATA6) and insulin receptor substrate proteins 2 (IRS-2).
CONCLUSIONSMiRNAs are differentially expressed between PCOS patients and controls. We identified and validated two miRNAs-miR-92a and miR-92b. They are significantly downregulated and may be involved in the pathogenesis of PCOS.
Adult ; Female ; Humans ; Hyperandrogenism ; genetics ; Insulin Resistance ; genetics ; physiology ; Male ; MicroRNAs ; genetics ; Ovary ; metabolism ; Polycystic Ovary Syndrome ; genetics
4.Synthesis of flavor esters catalyzed by CALB-displaying Pichia pastoris whole-cells in non-aqueous phase.
Zi JIN ; Ying LIN ; Dengfeng HUANG ; Guodong SU ; Shuangyan HAN
Chinese Journal of Biotechnology 2009;25(12):1927-1932
An enzyme-displaying yeast as a whole-cell biocatalyst seemed an alternative to immobilized enzyme, due to its low-cost preparation and simple recycle course. Here, we tried to use a recombinant Pichia pastoris displaying Candida antarctica lipase B (CALB) to catalyze the synthesis of short chain flavor esters in n-heptane. We studied some major influential factors of esterification reactions, such as carbon chain length of the substrates, alcohol structure, enzyme concentration, substrates concentration, molar ratio of the substrates. The acid conversions were determined by titration and gas chromatography analysis. About ten kinds of esters were synthesized successfully, and the acid conversions of eight esters reached as high as 90% after reaction for 6 h. The result also indicated that ethanol and hexanoic acid were the most suitable substrates for this whole-cell catalyst. Under the optimal reaction conditions (the amount of lipase 20 g/L (306.0 U/g-dry cell), hexanoic acid concentration 0.8 mol/L, the molar ratio of hexanoic acid to ethanol 1:1.1), hexanoic acid conversion reached 97.3% after reaction for 1.5 h. To our knowledge, the CALB-displaying P. pastoris whole-cell biocatalyst showed good tolerance for high substrates concentration and exhibited high reaction rate on esterification of short chain flavor esters among the present enzyme/cell reported. Thus, CALB-displaying P pastoris whole-cell biocatalyst was promising in commercial application for flavor esters synthesis in non-aqueous phase.
Biocatalysis
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Candida
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enzymology
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Enzymes, Immobilized
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Esters
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metabolism
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Fungal Proteins
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Lipase
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biosynthesis
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genetics
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Pichia
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genetics
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metabolism
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Recombinant Proteins
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biosynthesis
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genetics
5.Leukemia SH-1 cells purged by ZnPcH(1)-based photodynamic therapy.
Xiao-Lan LIN ; Hui-Fang HUANG ; Wan-Zi CHEN
Journal of Experimental Hematology 2012;20(4):842-846
The objective of this study was to investigate the effect of a novel Zinc phthalocyanine (ZnPcH(1)) based photodynamic therapy (PDT) on acute monocytic leukemia cell lines SHI-1 and its mechanism, so as to provide theory basis for bone marrow purging in vitro for patients with leukemia. The killing effect of ZnPcH(1)-PDT on SHI-1 cells were assessed by MTT method; the SHI-1 cell death patterns were analyzed by AO/EB fluorescence staining, TdT-mediated dUTP nick end labeling (TUNEL), DNA ploidy analysis, and Annexin V-FITC/PI double staining.Cell mixture was established by integrating SHI-1 cells with normal bone marrow MNC (by 1:100-1:10 000). Purging effect of ZnPcH(1)-PDT against SHI-1 mixed into normal MNC was assessed by analyzing the expression of fusion gene MLL/AF6 mRNA using nested RT-PCR. The results showed that ZnPcH(1)-PDT could effectively inhibit SHI-1 cell proliferation in dose-dependent manner, and ZnPcH(1)-PDT could induce cell apoptosis in time-dependent manner. 0.5 µmol/L ZnPcH(1)-PDT could completely photoinactivated kill SHI-1 cells in the simulated remission bone marrow. It concluded that ZnPcH(1)-PDT may be a effective and convenient promising purging technique for leukemia.
Apoptosis
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drug effects
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Bone Marrow Purging
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methods
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Cell Death
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drug effects
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Cell Line, Tumor
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Cell Proliferation
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drug effects
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Humans
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Indoles
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pharmacology
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therapeutic use
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Leukemia, Monocytic, Acute
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drug therapy
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pathology
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Organometallic Compounds
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pharmacology
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therapeutic use
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Photochemotherapy
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Photosensitizing Agents
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pharmacology
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therapeutic use
6.Changes of end-tidal carbon dioxide during cardiopulmonary resuscitation from ventricular fibrillation versus asphyxial cardiac arrest
Qing-Ming LIN ; Xiang-Shao FANG ; Li-Li ZHOU ; Yue FU ; Jun ZHU ; Zi-Tong HUANG
World Journal of Emergency Medicine 2014;5(2):116-121
BACKGROUND:Partial pressure of end-tidal carbon dioxide (PETCO2) has been used to monitor the effectiveness of precordial compression (PC) and regarded as a prognostic value of outcomes in cardiopulmonary resuscitation (CPR). This study was to investigate changes of PETCO2 during CPR in rats with ventricular fibrillation (VF) versus asphyxial cardiac arrest. METHODS:Sixty-two male Sprague-Dawley (SD) rats were randomly divided into an asphyxial group (n=32) and a VF group (n=30). PETCO2 was measured during CPR from a 6-minute period of VF or asphyxial cardiac arrest. RESULTS:The initial values of PETCO2 immediately after PC in the VF group were significantly lower than those in the asphyxial group (12.8±4.87 mmHg vs. 49.2±8.13 mmHg,P=0.000). In the VF group, the values of PETCO2 after 6 minutes of PC were significantly higher in rats with return of spontaneous circulation (ROSC), compared with those in rats without ROSC (16.5±3.07 mmHg vs. 13.2±2.62 mmHg,P=0.004). In the asphyxial group, the values of PETCO2 after 2 minutes of PC in rats with ROSC were significantly higher than those in rats without ROSC (20.8±3.24 mmHg vs. 13.9±1.50 mmHg,P=0.000). Receiver operator characteristic (ROC) curves of PETCO2 showed significant sensitivity and specificity for predicting ROSC in VF versus asphyxial cardiac arrest. CONCLUSIONS:The initial values of PETCO2 immediately after CPR may be helpful in differentiating the causes of cardiac arrest. Changes of PETCO2 during CPR can predict outcomes of CPR.
7.Immunological Reconstitution of T Cell Receptor Repertoire Diversity in Patients with Chronic Graft-versus-host Disease
Xin DU ; Yang-Qiu LI ; Wei LIN ; Jian-Hua SUN ; Zi-Lun HUANG ; Shu-Nong LI
Chinese Journal of Cancer 2001;20(1):32-34
Objective: The current stady was designed to investigate the reconstitution of T cell receptor repertoire in the patients with chronic Graft-versus-host disease (cGVHD). Methods: The TCR repertoire in peripheral blood mononuclear cells from 5 cGVHD patients was examined after PCR amplification of 24 Vβ gene subfamilies. Results: Only 2-8 Vβ subfamily T cells were found in the samples from these patients, and there were different demonstrations in different patients. We found Vβ T cells proliferated in 4 patients. Conclusion: The TCR repertoire complexities was abnormal in all patients,Vβ may be associated with cGVHD, and the method may be demonstrated the reconstitution of T cell after transplantation.
8.Study on Biological Characters of Sindbis Virus Yunnan Strain
Wen-li, HUANG ; Hai-lin, ZHANG ; Jing-lin, WANG ; Yu-zeng, ZHANG ; Zhu-qin, MI ; Hua-fang, SHI ; Deng-yun, ZI ; Guo-lin, ZHOU ; Guo-dong, LIAN
Virologica Sinica 2001;16(2):97-100
The biological characters of Sindbis virus strain of Yunnan(YN87448 strain) were studied by the test of the filtration, acid-resistant, ether-resistant, CPE, susceptibility of animal, HA, plague, determination of virus titres, and the cross-HI, cross-IFAT and PRNT as well.The results indicated that YN87448 strain belongs to Sindbis virus, Alphavirus, Togaviridae. YN87448 strain virus was plaque purified(PYN87448). The biological character of PYN87448 strain virus was studied too. PYN87448 strain virus will be used in the molecule biological test.
9.The role of peripheral blood mononuclear cells (PBMC) of HBV-infected mothers in the intrauterine infection of their fetuses.
Shu-hong LI ; Ya-fei YUE ; Shu-lin ZHANG ; Zi-yun SHI ; Qiao-di GUI
Chinese Journal of Hepatology 2006;14(4):264-267
OBJECTIVETo study the role of the HBV-infected mothers' PBMC in intrauterine transmission of HBV to their fetuses.
METHODSThirty pregnant women with serum HBV DNA negative and PBMC HBV DNA positive and their newborns were used as the study group. Ten pregnant women with serum HBV negative and their infants served as the control group. HBV DNA in serum and in PBMC was detected using nested polymerase chain reaction (n-PCR). The mothers' PBMC in newborns' peripheral blood was examined using heminested-PCR.
RESULTSFour newborns were serum HBV DNA positive and 8 newborns were HBV DNA positive in PBMC in the study group. Among them, 2 newborns were HBV DNA positive in both serum and PBMC, 6 cases were positive in PBMC only, and 2 cases were positive in serum only. Five mothers had the GSTM1 gene; and it was not detected in 3 newborns. Among the 8 newborns with HBV DNA positive in PBMC, 3 did not have the GSTM1 gene, at the same time their mothers possessed the GSTM1 gene. Mothers' PBMC were detected in all of these three newborns' peripheral blood. HBV DNA in serum and in PBMC of the control group infants were all negative.
CONCLUSIONHBV-infected PBMC of the mother may serve as a vector in HBV intrauterine infection.
Adult ; DNA, Viral ; analysis ; Female ; Hepatitis B virus ; isolation & purification ; Hepatitis B, Chronic ; blood ; transmission ; Humans ; Infant, Newborn ; Infectious Disease Transmission, Vertical ; Leukocytes, Mononuclear ; virology ; Pregnancy ; Pregnancy Complications, Infectious ; virology
10.Regulation of nitric oxide donor JS-K on tumor energy metabolism in H22 tumor-bearing mice
Ling LIU ; le Zi HUANG ; na Hui SHEN ; Jing WANG ; chen Mu LIU ; lin Chang HUA
Chinese Journal of Pharmacology and Toxicology 2017;31(7):730-735
OBJECTIVE To investigate the regulation of{O2 (2,4-dinitrophenyl)1-[(4-ethoxycarbonyl) piperazin-1-yl] diazen-1-ium-1, 2-diolate} (JS-K), a nitric oxide donor, on tumor energy metabolism in H22 tumor-bearing mice. METHODS The hepatoma animal model in BALB/c mice was established with H22 cell line. The inoculated mice were randomly divided into four groups. The JS-K group and model group received JS-K (0.75 and 1.50 mg?kg-1) and saline via tail the vein intravenously once every 3 d for 14 d, and 5 injections, respectively. The fluorouracil (5-FU) group received 5-FU 20 mg·kg-1 by intra-peritoneal injection once a day for 14 d. On the 15th day after the first administration, mice were sacri-ficed and the tumor, thymus and spleen were isolated and weighed immediately. The tumor growth inhibitory rate and organ index were calculated. The activities of hexokinase (HK), phosphofructokinase (PFK), pyruvate kinase (PK), succinate dehydrogenase (SDH), adenosinetriphosphatase (ATPase), and the levels of lactic acid (LD) and adenosine triphosphate (ATP) in tumor tissues were determined by colorimetric method. The expression of hypoxia-inducible factor 1 alpha (HIF-1α) and hexokinaseⅡ(HKⅡ) in the tumor tissue was analyzed by Western blotting. RESULTS Compared with model group, the tumor mass of JS-K 0.75 and 1.50 mg · kg-1 groups was significantly reduced (P<0.01), and the tumor growth inhibitory rate was 23.9%and 50.3%, respectively. There was no diffrence in thymic and splenic indexes between JS-K group and model group. The activity of HK, PFK, SDH, PK and ATPase of tumor tissue in model group was 22.6±3.7, 14.4±2.6, (10.5±2.6) U·g-1protein, (12.9±3.2) kU·g-1 protein and (0.70 ± 0.10) mmolPi · g-1protein · h-1, respectively, which dropped by 42.0%, 26.6%, 22.7%, 23.3%and 21.7%respectively (P<0.01, P<0.05) in JS-K 1.50 mg?kg-1 group. Compared with the model group, the level of ATP of tumor tissue in JS-K 1.50 mg?kg-1 groups dropped by 16.6%(P<0.01) and the level of LD in JS-K 0.75 and 1.50 mg?kg-1 groups dropped by 38.7%and 59.4%(P<0.01), respectively. In addi-tion, the expression of HIF-1αof tumor tissue in JS-K 1.50 mg?kg-1 group was decreased (P<0.01), and the expression of HKⅡ of tumor tissue in JS-K 0.75 and 1.50 mg?kg-1 groups was decreased signifi-cantly (P<0.05, P<0.01). CONCLUSION JS-K can inhibit the growth of tumor in H22 tumor-bearing mice and its mechanism may be related to regulating the tumor energy metabolism by inhibiting glycolysis and aerobic oxidation.