1.Comparison of effects of different anesthetic and postoperative analgesic methods on immune function of helper T lymphocytes and tumor metastasis after orthotopic liver cancer resection in rats
Di ZHOU ; Lanfang ZHU ; Changhong MIAO
Chinese Journal of Anesthesiology 2014;34(1):19-23
Objective To compare the effects of different anesthetic and postoperative analgesic methods on the immune function of helper T lymphocytes and tumor metastasis after orthotopic liver cancer resection in rats.Methods Orthotopic liver cancer was induced by intrahepatic tumor implantation (IHTI) with Morris hepatoma 3924A.Thirty male ACI rats receiving IHTI,aged 12-18 weeks,weighing 240-300 g,were randomized into 3 groups (n =10 each) using a random number table:control group (group C),general anesthesia combined with epidural block-postoperative epidural analgesia group (group GE + EA),and general anesthesia-postoperative intravenous analgesia group (group G + IA).The liver cancer resection was performed on 14th day after IHTI,group G + IA inhaled 5% sevoflurane for induction of anesthesia and inhaled 2%-3% sevoflurane and received intraperitoneal injection of morphine 20 μg/100 g for maintenance of anesthesia.In group GE + EA,general anesthesia was performed as the method previously described in group G + IA,and epidural block was performed with 0.25 % bupivacaine 25 μl after tracheal intubation.Within 3 days after operation,0.125 % bupivacaine 150 μl + morphine 20 μg were injected daily via an epidural catheter once every 4 h (25μl per time) for postoperative analgesia in group GE + EA,and postoperative analgesia was performed with intraperitoneal morphine 240 μg injected daily once every 4 h (40 μg per time) in group G+ IA.Before surgery (T1) and on 2nd,7th and 30th days after sugery (T2-4),blood samples were collected to detect the levels of plasma interferon-γ (IFN-γ),interleukin-4 (IL-4),IL-17,IL-10 and tumor growth factor-β1 (TGF-β1).IFN-γ/IL-4 ratio was calculated.The rats were sacrificed after collecting blood samples at T4,and the development of abdominal lymph node metastasis,malignant ascites,implantation metastasis to abdominal wall and visible pulmonary metastasis nodules were observed.Results Compared with C group,the incidence of pulmonary metastasis,abdominal lymph node metastasis,and malignant ascites was significantly decreased,the plasma IFN-γ and IL-17 levels at T2 and IL-4 and TGF-β at T2,3 were increased,and IFN-γ/IL-4 ratio was decreased at T2,3 in group G+ IA,and the incidence of pulmonary metastasis,abdominal lymph node metastasis,malignant ascites and implantation metastasis to abdominal wall was significantly decreased,the plasma levels of IFN-γ,IL-4,IL-17,IL-10 and TGF-β1 were increased at T2(P < 0.05),and no significant change was found in IFN-γ/IL-4 ratio in GE + EA group (P > 0.05).Compared with G + IA group,the incidence of pulmonary metastasis and abdominal lymph node metastasis was significantly decreased,the plasma levels of IFN-γ and IL-17 at T2 and IL-4 and IL-10 at T3 were decreased,the plasma levels of IFN-γ at T4 and TGF-β1 at T2 and IFN-γ/IL-4 ratio at T3,4 were increased in GE + EA group (P < 0.05).Conclusion The inhibitory degree of the immune function of helper T lymphocytes is decreased and development of tumor metastasis is reduced after orthotopic liver cancer resection in rats when general anesthesia combined with epidural block-postoperative epidural analgesia is applied as compared with those when general anesthesia-postoperative intravenous analgesia is applied.
2.Correlation between Toll-like receptor 4 single nucleotide polymorphism and sepsis in Chinese patients of Han nationality
Jing JIAO ; Lanfang ZHU ; Zhe LUO ; Changhong MIAO
Chinese Journal of Anesthesiology 2014;34(z1):62-66
Objective To investigate the correlation between Toll-like receptor 4 (TLR4) single nucleotide polymorphism (SNP) and the risk,severity and prognosis of sepsis in Chinese patients of Han nationality.Methods One hundred and three Han nationality patients who developed sepsis after surgery,aged 18-80 years,were enrolled in the sepsis group,and 114 Han nationality patients without sepsis after surgery,aged 18-80 years,were enrolled in the control group.Venous blood samples were taken from the peripheral vein and three SNPs in TLR4 gene,rs10759932,rs11536889 and rs2737190,were genotyped by matrix-assisted laser desorption ionization time of flight mass spectrometry analysis.Correction for Logistic regression analysis was made to eliminate the effects of sex,age,underlying diseases and operation methods.The correlation between genotypes of SNP and occurrence of sepsis,organ dysfunction,septic shock and death from sepsis was analyzed.The odds ratio (OR) and 95% confidence interval (Cl) were calculated.Results Compared with the control group,there was a significant difference in genotype frequency ratios of rs10759932 (P < 0.05),but there was no significant difference in genotype frequency ratios of the other two SNPs in sepsis group (P > 0.05).There was correlation between rs10759932 and the occurrence of sepsis,and the variant allele (CT + CC genotypes) of rs10759932 increased the risk of sepsis (OR =1.86,95% Cl 1.17-2.97,P < 0.05).There was no correlation between the three SNPs and sepsis-related organ dysfunction,septic shock and death from sepsis (P > 0.05).Conclusion There is correlation between the variant allele of TLR4 rs10759932 and the increase in risk of sepsis after surgery in Chinese patients of Han nationality.
3.Analysis of clinical and pathological characteristics of five cases with intravenous leiomyoma
Lanfang MIAO ; Ruixue LEI ; Yufen YUAN ; Peihong SHEN
Clinical Medicine of China 2016;32(1):61-64
Objective To investigate the clinical and pathological characteristics, diagnosis and differential diagnosis of uterine intravenous leiomyoma (IVL) , so as to improve the detection of the disease.Methods Five cases of IVL were retrospected, who were treated and diagnosed by clinical gross examination,morphologic features and immunohistochemical staining in Tumor Hospital of Anyang from 2011 to 2015.Results Symptoms of IVL were usually nonspecific, most patients experienced hypermenorrhea, abdominal pain or distension, and pelvic mass.Typical pathological changes were intravenous visible grey weblike or worm-formed nodules, which could be pulled out.Microscopically, the tumors were composed of mild spindle-shaped cells with rare mitotic figure and closely related to blood vessels.The tumors were positive for smooth muscle derived marker, progesterone receptor, estrogen receptor, and negative for S-100, HMB45 and CD34.However CD34 can displayed vascular endothelial cells around the tumor.Conclusion Uterine IVL is a benign tumor with malignant biological behavior, which has different clinical pathological features of leiomyoma ordinary.Surgery is the main treatment method.A satisfactory therapeutic effect can be achieved by selecting the appropriate extent of surgery.Long term follow up of iVL patients after operation is important.
4.Inhibition of bcl-2 gene expression by siRNA in HL-60 cells
Xiaoyong LEI ; Chunyan YAN ; Yulin TU ; Miao ZHONG ; Lanfang FENG ; Duanfang LIAO
Chinese Pharmacological Bulletin 2003;0(12):-
Aim To study the interference role of Bcl-2 siRNA on HL-60 cells. Methods Bcl-2 siRNA was synthesized in vitro transcription with silencer siRNA construction kit. The synthesized siRNA was transfected into HL-60 cells with lipid siPORT transfection. Forty-eight hours after the transfection, we used MTT and immunofluorescence to detect cell proliferation and apoptosis,and used RT-PCR and immunofluorescence to detect the level of Bcl-2 mRNA and Bcl-2 protein expression. Result Bcl-2 siRNA reduced the level of Bcl-2 mRNA and Bcl-2 protein expression in HL-60 cells and induced cell apoptosis. There was no difference on the effect of other groups compared with the control. Effective Bcl-2 siRNA specifically degraded Bcl-2 expression in the levels of mRNA and protein and induced HL-60 cells apoptosis.Conclusion These results indicate that siRNA is a highly specific tool for targeted gene knockdown. siRNA-mediated gene silencing is a reliable approach for large-scale screening of gene function and drug target validation.
5.Effect of 18alpha-glycyrrhizic acid and 18beta-glycyrrhizic acid on P-gp function and expression in Caco-2 cells.
Miao YAN ; Lanfang LI ; Huande LI ; Pingfei FANG ; Ping XU ; Mei ZHENG ; Danhua XU
China Journal of Chinese Materia Medica 2012;37(1):99-103
OBJECTIVEThe aim of the present study was to evaluate the modulating effect of glycyrrhizic acid C-18 epimers, 18alpha-glycyrrhizic acid (alpha-GL) and 18beta-glycyrrhizic acid (beta-GL) on both P-glycoprotein (P-gp) activity and expression in Caco-2 cell.
METHODThe effects of P-gp activity were analyzed by rhodamine (Rhd 123) accumulation test, and those of P-gp expression were analyzed by flow cytometry and real-time PCR.
RESULTAt middle and high concentrations (10, 60 micromol x L(-1)), alpha-GL inhibited the function of P-gp and with on dose dependent while beta-GL induced the function of P-gp at three test concentrations with no dose dependent too. At middle and high concentrations (10, 60 micromol x L(-1)), alpha-GL down-regulated the expression of MDR1 mRNA. At high concentrations (60 micromol x L(-1)), beta-GL up-regulated the expression of MDR1 mRNA; At high concentrations (60 micromol x L(-1)), beta-GL induced the expression of P-gp protein while alpha-GL has no effect on the expression of P-gp protein at three test concentrations.
CONCLUSIONThe effects of alpha-GL and beta-GL on the expression of MDR1 mRNA and CYP3A mRNA showed the same trend. The character that epimers of GL act on CYP3A and P-gp show similar stereo selectivity whether relate to PXR need further study.
ATP Binding Cassette Transporter, Sub-Family B ; genetics ; metabolism ; Caco-2 Cells ; Down-Regulation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression ; drug effects ; Glycyrrhizic Acid ; chemistry ; pharmacology ; Humans