1.Inhibitory effect of Saxagliptin on high glucose-induced overexpression of LncRNA-MALAT1 in endothelial cells
Xiaoyun HE ; Chunlin OU ; Yanhua XIAO ; Suxian ZHOU
Journal of Medical Postgraduates 2016;29(9):902-905
Objective Saxagliptin regulates the level of blood glucose by selectively inhibiting high-performance dipeptidyl peptidase 4, but its action mechanism is not yet clear .This study was to investigate the effect of the novel hypoglycemic agent Saxaglip -tin on the expression of long non-coding RNA (LncRNA) metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) and its target gene products transforming growth factor-β1 ( TGF-β1 ) in human umbilical vein endothelial cells ( HUVECs) stimulated by high glucose. Methods HUVECs were cultured in with D-glucose (D-GS) at the concentrations of 5.5, 10, 20, and 30 mmol/L and Saxagliptin at 0, 0.01, 0.1, 1, and 10μmol/L.The best concentrations of D-GS and Saxagliptin were determined as 30 mmol/L and 1 μmol/L, respectively.The HUVECs were divided into four groups:control (5.5 mmol/L D-GS), Saxagliptin (5.5 mmol/L D-GS+1 μmol/L Saxagliptin ) , high glucose ( 30 mmol/L D-GS ) , and high glucose +Saxagliptin (30 mmol/L D-GS +1μmol/L Saxaglip-tin), all cultured for 24 hours.Then the expressions of MALAT1 and TGF-β1 mRNA in the cells were detected by qRT-PCR, that of the TGF-β1 protein determined by Western blot , and the level of TGF-β1 in the supernatant measured by ELISA . Results The expressions of LncRNA-MALAT1 and TGF-β1 were significantly increased in the high glucose group as compared with the control ( 8.65 ±0.70 vs1.00 ±0.00 and 1.36 ±0.07 vs 1.00 ±0.00, P<0.01) but markedly inhibited in the high glucose +Saxagliptin group in compari-son with the high glucose group (2.17 ±0.24 vs 8.65 ±0.70 and 1.15 ±0.02 vs 1.36 ±0.07, P<0.05). Conclusion High glu-cose can induce the overexpression of LncRNA-MALAT1 and its target gene products TGF-β1 in HUVECs and cause damage to the cells, while Saxagliptin can significantly suppress this effect .
2.Expression of PirB in nerve tissues of normal adult mice
Xiaofeng CAI ; Rongdi YUAN ; Chunlin CHEN ; Yuanguo ZHOU ; Jian YE
Chinese Journal of Trauma 2011;27(9):844-847
ObjectiveTo investigate the expression of paired immunoglobulin-like receptor B (PirB) in optic nerve, visual cortex, cerebella, spinal cord and sciatic nerve of normal adult mice.MethodsTwelve healthy adult BALB/c mice were randomly and equally divided into two groups.The immunohistochemistry and Western blot were used to detect the expression of PirB in the tissues described above respectively.ResultsBoth the immunohistochemistry and Western blot test revealed that the expression of PirB was positive in the optic nerve, visual cortex, cerebella and spinal cord, but negative in the sciatic nerve.The positive signals in the sections were located in the cell bodies and the neurites were observed in some of them.Western blot showed the apparent positive band of PirB in the optic nerve, visual cortex, cerebella and spinal cord rather than in the sciatic nerve.The protein expression level of PirB was relatively high in the visual cortex (P <0.05) but relatively low in the optic nerves (P <0.01).ConclusionThe PirB expresses positively in the optic nerve, indicating that PirB protein may closely correlate with the poor regeneration of the optic nerve.
3.Comparative study on satisfaction of young medical workers in Shanghai
Meng WANG ; Hong LIU ; Lan ZHOU ; Keqin RAO ; Chunlin JIN
Chinese Journal of Hospital Administration 2016;32(10):797-799
Objective To find out the satisfaction changes among young medical workers and explore the influence of the healthcare reform on such people.Methods A survey was conducted by means of random sampling among 716 medical workers under 45 years old from 6 hospitals,and combined with the survey data in 2009,young medical workers′satisfaction changes and the influencing factors were analyzed.Results Young medical workers′satisfaction is generally low,with declining satisfaction of social respect as well;satisfaction of medical workers in community medical institutions and secondary medical institutions was found declining dramatically.Conclusions Healthcare reform should pay more attention to these young people in terms of their own satisfaction,especially for those working at primary healthcare institutions.
4.Expression of Th1/Th2 Cytokines in Early Syphilitic Lesions and Its Relationship with the Sero-conversion of RPR Test in Patients with Syphilis
Pingyu ZHOU ; Sidong XIONG ; Chunlin YAN ; Jinhua XU ; Kanghuang LIAO
Chinese Journal of Dermatology 2003;0(12):-
Objective To investigate the expression of Th1/Th2 cytoki nes in early symptomatic syphilitic lesions and its relationship with the sero-c onversion of RPR test in patients with syphilis. Methods The expression of Th1/Th2 cytokines in lesions from 30 patients with early symptomatic syphilis was i mmunohistochemically detected with ABC method. The serum titers of RPR test in t hese patients were measured in 1, 3, 6, 9 and 12 months after routine benzathine penicillin treatment. Results Among 10 cases of primary syphilis, the express ion of both Th1 and Th2 cytokines in the lesions was found in 9 cases, while onl y Th1 cytokine expression was observed in the remainder case; and the sero-conve rsion of RPR test occurred in all 10 cases during the follow-up period. Among 20 cases of secondary syphilis, Th1/Th2 cytokines expressed in the lesions in 16 c ases, and only Th2 cytokines expressed in 4 cases; and the sero-conversion of RP R test was found in 12 cases during the follow-up period. The expression of Th1 cytokines in early syphilitic lesions was positively correlated with the sero-co nversion of RPR test. The higher the expression of IFN- the more likely the s ero-conversion of RPR test. Conclusion The early activation and persistence of the expression of Th1 cytokines may play an important role in the clearance of pathogens.
5.Distribution and drug resistance of pathogens for bacterial infection after lung transplantation
Chunlin ZHANG ; Lingling LI ; Jian ZOU ; Jingyu CHEN ; Ying YIN ; Min ZHOU ; Weizhen QIAO
Chinese Journal of Organ Transplantation 2016;37(2):95-100
Objective To analyze the distribution and drug resistance of pathogens for bacterial infection after lung transplantation,so as to provide evidence for clinical prophylactic strategies postoperation and reasonable use of antibiotics.Method The bacterial distribution and drug resistance of 81 recipients after lung transplantation in our hospital were retrospectively analyzed from May 2009 to October 2012.The VITEK-32 full-automatic microbial identification system (Biomerieux,France)and its supplementary reagent were used for bacterial identification and drug sensitive test.The data were statistically analyzed by using the software SPSS 13.0.Result There were 67 cases of bacterial infection in the 81 recipients after lung transplantation and the infection rate was 82.72% (67/81).The infection was caused by one kind of bacteria in 20 patients,two kinds of bacteria in 23 patients and multiple bacteria in 24 patients.157 strains pathogenic bacteria were produced,and the grampositive bacilli and the gram-negative bacilli accounted for 12.74% and 87.26% respectively.The most common pathogens for the bacterial infection were Acinetobacter baumannii,Klebsiella pneumoniae,Pseudomonas aeruginosa,Stenotrophomonasmaltophilia,Escherichia coli and Staphylococcus aureus.Most of the bacterial infections occurred in the early period (≤1 month) after lung transplantation and most non-fermentative bacterial pathogens were resistant to multi-antibiotics.Conclusion The bacterial infection rate is high after lung transplantation.The rational use of antibiotics in clinical practice should be adjusted according to the bacterial distribution and drug resistance.
6.Spectroscopic Study of Interaction of Harpagoside and Human Serum Albumin
Tuanwu CAO ; Kun ZHOU ; Wenbing HUANG ; Jianwei SHI ; Xiaoping TAN ; Chunlin HUANG ; Ai RAN
Chinese Journal of Analytical Chemistry 2017;45(5):700-706
Harpagoside (HAR) is believed to be a main compound in Scrophularia ningpoensis which possess a broad of biological activities.Human serum albumin (HSA) has important physiological roles in transportation, distribution and metabolism of many endogenous and exogenous substances in body.It is great significance in pharmacology to investigate the interaction mechanism of HAR and HSA.In this work, the interaction between HAR and HSA was investigated by fluorescence and ultraviolet absorption spectroscopy at different pH (pH=4.0, 7.4, and 9.0) and temperatures (297, 310 and 323 K).The experimental results showed that the HAR could cause the fluorescence quenching of HSA through a static quenching procedure, showing that the HAR regularly quenched the intrinsic fluorescence of HSA, and a decrease in the quenching constant was observed with an increase in temperature.Under different conditions, all the magnitude of binding constants (KA) was larger than 105 L/mol and the number of binding sites (n) in the binary system were approximate to 1.Base on the magnitude of enthalpy and entropy changes, the negative values of ΔG, ΔH and ΔS revealed that the binding of HAR with HSA was spontaneous and exothermic process, and the main interaction forces of the HAR with HAR were van der Waals forces and/or hydrogen bonding interaction.The binding distance (r) between the HAR and HSA was calculated to be about 4.2 nm based on the theory of F(o)rster′s nonradiation energy transfer, which indicated that the energy transfer from HSA to HAR occurred with high possibility.What was more, the synchronous florescence spectroscopy confirmed the conformational changes of HSA during the binding reaction.
7.Mitochondrial DNA 8 point mutations in patients with type II diabetes mellitus
Songmei LIU ; Xin ZHOU ; Xia LI ; Fang ZHENG ; Han QIN ; Chunlin CAI
Chinese Journal of Pathophysiology 1986;0(03):-
AIM: To explore the relationship between various mitochondrial (mt) DNA tRNA Leu (UUR) and ND1 gene mutations and type 2 diabetes mellitus (T2DM) among Chinese in Hubei Province. METHODS: PCR restriction fragment length polymorphism (PCR-RFLP) analysis was used to screen point mutations of mtDNA ( 3 243, 3 256, 3 290, 3 316, 3 394, 3 421, 3 426, 3 460, 3 593) in 174 T2DM and 207 healthy controls. Then, DNA sequencing, reverse dot blot hybridization and Genchip were used to compare and confirm mutations. All mutations were analyzed by DNASTAR and Antherprot softwares. RESULTS: In diabetic group, there were 5 carriers (2.9%) of 3 316 G→A (Ala→Thr) mutation, 4 (2.3%) of 3 394 T→C (Tyr→His) mutation, 1 (0.6%) of 3 593 T→C(Val→Ala) mutation, and 1 (0.6%) of 3 618 T→C(Phe→Phe) mutation. Among 3 316 (G→A) mutations , there were more than 1 point mutations in 2 cases, one accompanied with 3 256 C→T(Arg→Arg) and 3 688 G→C (Ala→Pro) mutations, another accompanied with 3 606 A→G(Leu→Leu) mutation. 3 606 (A→G), 3 618 (T→C) and 3 688 (G→C) were novel mutations, GenBank accession number is DQ092356. In controls, only 3 316 (G→A) mutation was found in 1 subject (0.5%). There was significant difference between two groups for 3 394 (T→C) mutation frequencies (P
8.ASSAYING OF ARSENIC CONTENT IN SOIL SAMPLES FROM FIELD USED MAGNESIUM BORATE ORE POWDER AS MOLLUSCICIDE
Tao ZHANG ; Yixin HUANG ; Yun ZHOU ; Lixing ZHENG ; Chunlin ZHAO ; Qingw JIANG
Chinese Journal of Schistosomiasis Control 1989;0(03):-
Objective To evaluate the safety of magnesium borate ore powder (MBOP) as a molluscicide, especially for the arsenic content in the soil samples from the fields used MBOP over more than one year. Method The arsenic in the soil samples was assayed by using the AgDDTC method recommended by the government. Result In all of the soil samples, the arsenic contents did not exceed the national health criterion. However, the arsenic in the MBOP treated soil samples was higher than that in the control one′s. Conclusion The application of MBOP as a molluscicide in the fields does cause some side effects on the environment.
9.Three Scutellaria baicalensis extracts induce the apoptosis of a human cutaneous squamous cell carcinoma cell line SCL-1: an experimental study
Mei LIU ; Chunlin ZHOU ; Hong ZHU ; Guangyu JIN ; Hongduo CHEN ; Chundi HE
Chinese Journal of Dermatology 2014;47(9):650-653
Objective To investigate the effect of three Scutellaria baicalensis extracts (baicalin,baicalein and wogonin) on the apoptosis of a human cutaneous squamous cell carcinoma cell line,SCL-1.Methods Methyl thiazolyl tetrazolium (MTT) assay was performed to detect the proliferation of cultured SCL-1 cells treated with different concentrations (12.5,25,50,75 and 100 μmol/L) of baicalin,baicalein and wogonin for various durations (12,24 and 48 hours).Some SCL-1 cells were treated with baicalin,baicalein and wogonin of 12.5 μ mol/L respectively for 48 hours followed by the detection of cell apoptosis by double staining with annexin V-fluorescein isothiocyanate/propidium iodide in combination with enzyme-linked immunosorbent assay (ELISA),as well as estimation of cell cycle by flow cytometry.The 50% inhibitory concentration was determined by line regression model and inner insert method,and statistical analysis was carried out by Student's t test,one-way analysis of variance (ANOVA),and Student-Newman-Keuls-q test.Results Baicalin,baicalein and wogonin all inhibited the proliferation of SCL-1 cells in a dose-and time-dependent manner (all P < 0.01).Under the same conditions (treatment concentration and duration),baicalein showed the strongest inhibitory effect on the proliferation of SCL-1 cells,followed by wogonin and baicalin (all P < 0.01).All the Scutellaria baicalensis extracts induced the apoptosis of SCL-1 cells and arrested them in G1-phase.The percentage of cells in G1 phase was 56.37% ± 2.41%,74.23% ± 2.02% and 64.15% ± 1.87%,and early apoptosis rate was 8.09% ± 1.02%,24.13% ± 0.76% and 14.45% ± 1.57%,in SCL-1 cells treated with baicalin,baicalein and wogonin of 12.5 μmol/L for 48 hours,respectively,compared to 45.04% ± 1.93% and 4.12% ± 0.29% in the untreated control cells respectively (F =83.29,186.37,respectively,both P < 0.01).Similarly,there was a downward trend from baicalein to wogonin and baicalin in the effect on cell apoptosis and cell cycle arrest of SCL-1 cells.Conclusions Scutellaria baicalensis can inhibit the growth and induce the apoptosis of SCL-1 cells,which may provide new ideas for the treatment of cutaneous squamous cell carcinoma with traditional Chinese drugs.
10.Effect of stromal cell derived factor-1αand interleukin-1βon inducing vascular endothelial cells expressing lymphatic phenotype
Ning SUO ; Xueying WANG ; Chunlin YANG ; Hui ZHOU ; Fei LI ; Zongpu ZHANG ; Fangzhu WAN ; Hua TIAN
Acta Anatomica Sinica 2014;(3):388-392
Objective To investigate the effect of stromal cell-derived factor-1α( SDF-1α) and interleukin ( IL-1β) on inducing vascular endothelial cells to express lymphatic phenotype .Methods The CRL-1730 cell line was cultured and treated with SDF-1αor IL-1β.The expression of endothelial cell markers and lymphatic endothelial cell markers were investigated with Real-time PCR, Western blotting and immunocytochemistry .Results In CRL-1730 cell line, endothelial cell markers such as voln willebrand factor ( vWF ) , VE-cadherin , vascular endothelial growth factor receptor(VEGFR)2, were dose dependently down-regulated after SDF-1αstimulation, while lymphatic phenotypes such as Prox-1, podoplanin and lymphatic vessel endothelial hyaluronan receptor-1(LYVE-1), were dose-dependently up-regulated after SDF-1αstimulation.The changes of vWF, VEGFR2 and podoplanin, Prox-1, LYVE-1 expression after IL-1βstimulation was similar to that after SDF-1αwhile expression of VE-cadherin changed slightly .Conclusion SDF-1αand IL-1βare able to induce vascular endothelial cell expressing lymphatic phenotype .