1.Association between vitamin D level and bone mineral density in aged people
Zhili GAO ; Xiaohong DU ; Zaisheng ZHU
Chinese Journal of Geriatrics 2012;31(6):513-515
Objective To study the association between vitamin D status and bone mineral density(BMD) in aged people.Methods A total of 118 patients admitted in cadre department aged (77.7±11.2) years were selected from July 2010 to May 2011.The BMD at lumbar spine (L1-4) and femoral neck was measured by dual energy X-ray absorptiometry (DEXA).According to BMD,patients were divided into two groups:osteoporosis and non-osteoporosis group.The data of serum 25-hydroxy vitamin D3 [25(OH)D3]concentration,height,weight and age of patients were collected at the same time.Results The age and body mass index (BMI) in osteoporosis group were (81.6 ±5.6) years and (22.5 ± 4.0 ) kg/m2,while (79.4 ± 6.9 ) years and (24.1± 4.2 ) kg/m2 in nonosteoporosis group (t=1.80 and -2.01,P>0.05).The concentrations of serum 25(OH)D3 in two groups were(21.6± 10.3)nmol/L and (32.0± 13.8) nmol/L,respectively(t=-4.20,P<0.01).And there were 95.3 % (41/43 )and 81.3 % (61/75) of patients whose serum 25 (OH)D3 level were ≤50nmol/L in osteoporosis and non- osteoporosis group,respectively(x2 =4.58,P<0.05).Furthermore,the 25 (OH) D3 level was positively correlated with BMD at femoral neck(r=0.22,P<0.05),but not correlated with BMD at L1-4 ( r=0.18,P>0.05).Conclusions Vitamin D status is correlated with BMD at femoral neck in aged people.
2.Microarray analysis of differentially gene expression profile in LPS-stimulated primary Kupffer cells
Zhili TAN ; Tong ZHU ; Wenjuan TU ; Liangming LIU
Chinese Journal of Immunology 2016;32(12):1734-1740
Objective:To investigate the changes of gene expression profile in lipopolysaccharide (LPS)-stimulated primary Kupffer cells ( KCs ) . Methods: Rat KCs were isolated and purified by means of in situ perfusion and density gradient centrifugation. After being identified by ink phagocytosis and ED2 staining test,KCs were stimulated with LPS. Gene expression profile were studied using gene microarrays,and the most significant upregulated gene was verified using real-time PCR. Results:27 genes were upregulated including Ces1f, Slc17a3, Slc21a4, Hsd17b2, Sorbs2, Ccdc116, Mgam, Myo5b, Etl4, Fabp1, Kif4b, Fosl1, Cyp4a1, Penk, Tmem221,Rpl5,Nr2f1,Hoxb1,Gpr165,Fam90a13p,Kpna6,Irak1bp1,Kcnh1 and 4 unnamed genes and 4 downregulated including Oc90,Tagln,Arxes2 and Olr830 in LPS-stimulated KCs. Among the upregulated genes, Ces1f was the most significant upregulatory gene. Real-time PCR confirmed that the levels of Ces1f were 23. 88 times higher in LPS-stimulated than control cells. Conclusion:There is a significant difference between LPS-stimulated and normal control cells in gene expression profile by microarray analysis,and Ces1f is the most significantly upregulated gene.
3.Effects of inhibited expression of IRF3 in LPS-stimulated Kupffer cells on the activation of signal transduction pathways
Tong ZHU ; Wenjuan TU ; Zhili TAN ; Liangming LIU
Chinese Journal of Microbiology and Immunology 2015;(8):587-594
Objective To investigate the effects of interferon regulator factor 3 (IRF3) shRNA on the expression of TLR4 downstream signal molecules including IRF3-IFN-β, NF-κB/p38 MAPK-TNF-α/IL-1βand IL-10 in lipopolysaccharide (LPS)-stimulated Kupffer cells (KCs). Methods KCs were isolated from rats by in situ perfusion. The adenovirus strains carrying IRF3 shRNA were used for the transfection of purified KCs. The isolated KCs were randomly divided into four groups including adenovirus(-) LPS(-) treatment group, adenovirus(-) LPS(+) treatment group, adenovirus(+) LPS(-) treatment group and ad-enovirus(+) LPS(+) treatment group. The levels of cytokines in the supernatants of KC culture were detec-ted by enzyme-linked immunosorbent assay ( ELISA ) . Real-time PCR and Western blot assay were per-formed to analyze the expression of related cytokines at mRNA and protein levels, respectively. Results The expression of IRF3 at mRNA and protein levels in primary cultured KCs were induced by LPS. The cel-lular constitutive expression of IRF3 at mRNA level and the LPS-induced expression of IRF3 were signifi-cantly inhibited after transfection of KCs with adenovirus strains carrying IRF3 shRNA. However, the nucle-ar constitutive expression of IRF3 protein was not affected by IRF3 shRNA. The expression of IFN-βat mR-NA and protein levels in KCs were induced by LPS, but were suppressed by the interference with IRF3 shR-NA. No significant changes of the cellular constitutive expression of IFN-βat mRNA and protein levels were observed in IRF3 shRNA-treated KCs. Enhanced expression of proinflammatory cytokines including TNF-αand IL-1β at mRNA and protein levels were detected in LPS-stimulated KCs. Transfection of KCs with ade-novirus strains carrying IRF3 shRNA inhibited the LPS-induced secretion of TNF-α and IL-1β, but neither LPS-induced expression of TNF-α and IL-1β at mRNA level nor cellular constitutive expression of TNF-αand IL-1βat mRNA and protein levels were affected by IRF3 shRNA. The LPS-induced expression of IL-10 at mRNA and protein levels were enhanced in IRF3 shRNA-treated KCs. However, the cellular constitutive expression of IL-10 at mRNA and protein levels were not affected by the adenovirus. The levels of phosphor-NF-κB p65 subunit and phosphor-p38 MAPK protein in the nuclei of KCs were increased upon the stimula-tion with LPS. Treatment of KCs with IRF3 shRNA showed no significant effects on nuclear phosphor-NF-κB p65 subunit and phosphor-p38 MAPK. Conclusion Transfection of LPS-stimulated primary KCs with ade-novirus strains carrying IRF3 shRNA could effectively inhibit the expression of IRF3 and the transduction of downstream signals. IRF3 enhanced the secretion of TNF-αand IL-1β, but inhibited the expression of IL-10 in LPS-treated KCs. The LPS-induced activation of NF-κB and p38 MAPK in KCs were not affected by IRF3 signal.
4.A cumulative Meta analysis of association between H .Pylori infection and pancreatic cancer
Yong DENG ; Weiqiong ZENG ; Zhu ZHAN ; Zhili LIU
Chongqing Medicine 2015;(32):4549-4552
Objective To obtain a reliable estimate of the risk of H. pylori infection in causing pancreatic cancer ,by perform‐ing a M eta‐analysis of the existing observational studies evaluating the association .Methods Observational studies comparing the prevalence of H. pylori infection in patients with pancreatic cancer and healthy controls were identified through systematic search in the Medline ,EMBASE ,the Cochrane ,PubMed ,VIP database .H. pylori infection was confirmed by serological testing using an anti‐gen‐specific enzyme‐linked immunosorbent assay .Pooled adjusted odds ratios (AOR) and associated 95% confidence intervals (CI) were obtained by using a Dersimonian and Laird random‐effects model .Results Six studies involving a total of 2 335 patients met our eligibility criteria .A significant association between H. pylori seropositivity and development of pancreatic cancer (AOR=1 .38 , 95% CI:1 .08-1 .75 ;P=0 .009) was seen .No significant association had been seen on pooled analysis of the three studies assessing the relationship between CagA positivity and pancreatic cancer (AOR=1 .14 ,95% CI:0 .66-1 .97 ,P=0 .639) .Conclusion The da‐ta suggests an association between infection with H. pylori and the development of pancreatic cancer .Further research is needed to confirm our findings .
5.Impacts of adenovirus on the transfection efficiency and proliferative activity of primary Kupffer cells
Tong ZHU ; Wenjuan TU ; Zhili TAN ; Liangming LIU
The Journal of Practical Medicine 2016;32(6):879-882
Objective To investigate the impacts of adenovirus on the transfection efficiency and proliferative activity of primary Kupffer cells (KCs). Methods Rat liver KCs were separated and purified by density gradient centrifugation , and was then transfected with adenovirus carrying green fluorescence protein (GFP) gene at different multiplicity of infection (MOI). After 24 h, the transfection efficiency was evaluated by fluorescence microscope and flow cytometry. The proliferative activity of KCs was assessed by colorimetric method. Results The positive percentages of GFP staining cells were statistically different among different doses of adenovirus (MOI 0, 100, 300, 500, 700 and 900) under fluorescence microscopy or by flow cytometry (P <0.05 for all comparisons). The cell proliferative activity had significant differences among MOI 300, 500, 700 and 900(P < 0.05 for all comparisons), but had no differences among MOI 0, 100 and 300 (P > 0.05 for all comparisons) by CCK8 assay. Conclusions KCs can effectively be transfected by GFP adenovirus; and with an increase in virus MOI, the transfection efficiency rises gradually. A higher dose of adenovirus may have a negative effect on cell proliferative.
6.EFFECTS OF COMBINED ANTIOXIDANTS ON THE EXPRESSION OF CRP AND IL-6 IN TYPE 2 DIABETIC PATIENTS
Songchao GUO ; Cuifeng ZHU ; Zhili ZENG ; Qingxiang WU ; Zuojie LUO
Acta Nutrimenta Sinica 2004;0(05):-
Objective: To investigate the effects of the cooperative actions of isoflavone, vitamin C and E on the expression of acute C reactive protein (CRP) and interleukin 6 (IL-6) in the patients with typeⅡdiabetic mellitus (DM). Method: Ninety six patients with DM were divided randomly into 4 groups, DM control group(B), and groups receiving low(C), medium (D) , and high (E) dose of the compound antioxidant of isoflavone, vitamin C and E respectively. Another 24 healthy subjects served as normal control group (A). Observe the changes of the levels of CRP and IL-6 during oral glucose tolerance test (OGTT). Results: The peak of serum levels of CRP and IL-6 in the patients of DM control group and lower dosage group appeared 1 h after OGTT and were significantly higher than the 0 h value (P
7.Effects of IRF3 gene interference on dynamic changes of cytokine secretions in early of LPS stimulation in primary Kupffer cells
Tong ZHU ; Wenjuan TU ; Zhili TAN ; Liangming LIU
Chinese Journal of Immunology 2016;32(4):470-475,479
Objective:To investigate the effects of IRF 3 shRNA adenovirus on dynamic changes of early cytokines in LPS-stimulated primary Kupffer cells ( KCs ).Methods: Rat KCs were isolated and purified by means of in situ perfusion.After being infected with adenovirus carrying IRF 3 shRNA for 48 h, KCs were stimulated with LPS.Cell culture supernatants were collected respectively at 0,2,4 and 6 h after LPS stimulation as well as cells at 6 h.Supernatant cytokine secretion levels were detected by enzyme-linked immunosorbent assay ( ELISA).Intracellular gene expressions were tested by RT-PCR and Westeron blot.Results:IRF3 mRNA and protein were induced by LPS ,but suppressed by IRF 3 shRNA adenovirus in LPS-stimulated or non-stimulated KCs.IFN-βsecretions rose in the very early stage ( at 2 h) ,reached the peak at 4 h,and began to reduce but still remained high levels at 6 h after LPS stimulation in KCs.Interference adenovirus pretreatment suppressed IFN-βsecretions ( especially the secretion peak ) at each time point after LPS stimulation.IFN-βsecretions reached normal levels at 6 h after the stimulation in adenovirus-pretreated cells;TNF-αse-cretions rapidly increased in the very early stage and reached the peak at 2 h,then began to decrease gradually ,but remained high levels at 6 h after LPS stimulation in KCs.Interference adenovirus pretreatment inhibited LPS-induced TNF-αsecretions, especially the secretion peak;IL-1βsecretions did not increase untill 4 h, but reached a higher level at 6 h after LPS stimulation.Interference adenovirus suppressed IL-1βsecretions in the early stage of LPS stimulation;IL-10 secretions began to rise in the very early stage ,and gradually increased over time after LPS stimulation in KCs.Pretreatment of adenovirus with IRF 3 shRNA promoted upregulations of IL-10 secretions at each time point of the early of LPS stimulation.Conclusion:IRF3 gene expression can be silenced by IRF 3 shRNA ad-enovirus.IRF3 can promote its downstream signaling molecule IFN-βand pro-inflammatory cytokines including TNF-αand IL-1β,and block anti-inflammation cytokine IL-10 secretions in LPS-stimulated primary KCs.Therefore,IRF3 may play a central role in immune inflammatory injury of liver tissues.
8.The Inhibitory Effect of Down-Regulating Phosphorylated HER2 on Proliferation and Metastasis in U2-OS Osteosarcoma Cells
Zhihong ZHANG ; Xinhua LONG ; Zhili LIU ; Heng WANG ; Taofang WANG ; Liangbo ZHU
Tianjin Medical Journal 2014;(1):1-3,4
Objective To investigate the effects of down-regulating phosphorylated human epidermal growth factor receptor 2 (HER2) on the proliferation and metastasis in human osteosarcoma cells (U2-OS) in vitro. Methods Various concentrations of HER2 phosphorylation inhibitor lapatinib ditosylate (5, 10, 20, 30 and 40 μmol/L) were adopted to deal with U2-OS. MTT assay was performed to evaluate the cell proliferation during various times (24, 48 and 72 h), and the IC50 value in 24 h was calculated. The value of 10μmol/L (IC50=22.15μmol/L) was chosen to deal with U2-OS cells. The expres-sion level of phosphorylated HER2 (p-HER2) was measured by Western blot assay. The cell migration and invasion abilities were detected by Wound healing and Transwell invasion assays. Results The cell proliferation of U2-OS was significantly inhibited by HER2 phosphorylation inhibitor lapatinib ditosylate in a concentration- and time-dependent manner. During 24 hours, the p-HER2 level was significantly lower in lapatinib ditosylate group than that of negative control group (0.093± 0.033 vs 0.306±0.033), the cell migration rate was significantly lower in lapatinib ditosylate group than that of negative con-trol group (32.70%±3.00%and 94.52%±4.76%), and the trans-membrane cells were significantly lower than those of nega-tive control group (37/HP±5/HP and 85/HP±10/HP), respectively. Conclusion The down-regulating p-HER2 in U2-OS could efficiently inhibit the cell proliferation, migration and invasion in vitro. HER2 has the potential to become a molecular target for anti-osteosarcoma metastasis.
9.Cultivation of doctor-patient communication ability for general practitioners based on iceberg theory
Xiayuan ZHU ; Hao WU ; Caiying GE ; Xiaoxia LI ; Zhili LI ; Min KONG ; Xiumei LIU ; Junxing LIU
Chinese Journal of General Practitioners 2017;16(9):737-739
An iceberg model for better understanding the patients was developed.In this model the seeking for consultation is visible part of illness behavior,but the thoughts,concerns and expectations toward his or her illness are the hidden parts.Through detailed history taking,using appropriate language skill (Leichester Assessment Package) and nonverbal communication skill (space distance,eye contact,listening,pronunciation,appropriate silence),physicians can acquaint comprehensive knowledge about the hidden thoughts,concern and expectation of patients about their illness.This model would help general practitioners to raise communication and consultation ability and skills in their practice.
10.Research capacity training for graduate students in basic medical sciences with the guideline of translational medicine
Jian ZHOU ; Hongmei XU ; Xiaoyun SUN ; Zhili ZHU ; Yuzhang WU ; Jijian ZHANG
Chinese Journal of Medical Science Research Management 2015;28(3):250-252
In China,few research findings in basic medicine could be translated to clinical practice,which is known as the disconnection between basic research and clinical application.Such phenomenon is mainly due to lack of ability in translational medicine,particularly,lack of training in clinical research and translational medicine.Now the emergence of translational medicine has put forward new requirements for graduate education in medical school,and demanding that the patient-centered concept be strengthened.As the major force of scientific and technological innovation,graduate students should receive long-term systematic trainings and conduct original researches.In addition,advanced courses in clinical research and translational medicine should be offered to graduate students ;furthermore,translation research platforms should be built up for them so as to improve the capacity of scientific research and innovation.