1.1,25-Dihydroxyvitamin D3 and Its Analogues Modulate the Expression and Secretion of IL-10 by Human Monocyte-derived Dendritic Cells
Kejian ZHU ; Limin LAO ; Xiaohong MAO
Chinese Journal of Dermatology 2003;0(09):-
Objective To elucidate the therapeutic mechanism of 1,25-dihydroxyvitamin D3 and its analogues in psoriasis. Methods The modulating role of calcitriol, calcipotriol and tacalcitol on the expression and secretion of IL-10 in monocyte-derived dendritic cells (MDDCs) was investigated. Human monocytes were differentiated into MDDCs in vitro in the presence of GM-CSF and IL-4. Semi-quantitative RT-PCR and ELISA were used to analyze the mRNA expression and protein production of IL-10 in MDDCs. In each experi-ment, 24,25-dihydroxyvitamin D3 (a vitamin D3 metabolite with a low affinity to the vitamin D receptor) was used as a control. Results The mRNA expression and protein secretion of IL-10 in MDDCs was up-regulated by calcitriol, calcipotriol or tacalcitol with a similar potency, but not by 24,25-dihydroxyvitamin D3. The up-regulatory role of calcitriol on IL-10 protein secretion was dose dependent. Conclusion The therapeutic effect of calcitriol and its analogues in the treatment of psoriasis may be related to the induction of IL-10 production.
2.Internalization and down-regulation of the epidermal growth factor receptor in a human keratinocyte cell line HaCaT
Xiaohong MAO ; Fengzhi XU ; Lunfei LIU ; Jisu CHEN ; Limin LAO
Chinese Journal of Dermatology 2008;41(12):814-817
Objective To understand the molecular mechanism underlying the epidermal growth factor receptors(EGFR)signal transduction and its feed-back regulation.Methods Two human keratinocyte cell lines,HaCaT and CHOwt,were cultured and treated with a certain concentration of different ligands,including epidermal growth factor(EGF),heparin-bounding(HB)-EGF,transforming growth factor α (TGFα)and heregulin(HER),for various durations(2,4,8,16,20 hours).After the treatment,cells were collected and protein was extracted.The amount of total and active EGFR was measured by immunoprecipitation and immunoblot assay.The internalization and down-regulation of EGFR were visualized with immunofluorescence and laser seanning confocal microscopy.Results As shown by immunoblot technique,EGF and HB-EGF continuously down-regulated the total amount of EGFRs,whereas TGFα and HER had no significant effect on the degradation of EGFRs.The activation of EGFRs was also attenuated to different extent after long-time treatment with EGF,HB-EGF and TGFα.As indirect immunofluorescenee revealed,in untreated HaCaT and CHOwt cells,EGFRs were essentially located at the plasma membrane,with a little cytosolic distribution;after ten-minute treatment with EGF,EGFRs clustered into patch-like structures which were particularly obvious in HaCaT cells,and translocated into cytoplasmic vesicles resembling endosomes (relatively apparent in CHOwt cells),while the total amount of EGFRs remained constant in these cells.The fluorescence signal from the total EGFRs decreased evidently after four-hour treatment with EGF,indicating a strong reduction in the receptors.Conclusions EGF and HB-EGF,but not TGFα or Heregulin,could down-regulate the amount of total and active EGFRs.There might be different mechanisms for the signal transduction related to EGFRs intemalization and down-regulation between HaCaT and CHOwt cells.
3.Expression of IFN-? and IL-18 in Peripheral Blood of Psoriasis
Limin XU ; Yanhong WANG ; Hong LI ; Shuhe MAO
Chinese Journal of Dermatology 2003;0(10):-
Objective To investigate the expression of IFN-? and IL-18 in patients with progressive psoriasis and its clinical significance. Methods Twenty cases of psoriasis vulgaris, 19 cases of erythroder-mic psoriasis and 10 oases of pustular psoriasis were studied. Sandwich ELISA was used to detect the plasma level of IFN-? and IL-18, or supernatant level of IFN-? and IL-18 in PBMC culture under the stimula-tion of staphylococcal enterotoxin B (SEB). Results The plasma level of IFN-? in patients with psoriasis vulgaris, erythrodermic psoriasis and pustular psoriasis was higher than that in healthy control. The plasma level of IL-18 in progressive psoriasis vulgaris was higher than that in healthy control. The supernatant level of IFN-? and IL-18 in PBMC culture in progressive psoriasis vulgaris were higher than that in normal control, with a positive correlation between IFN-? level and IL-18 level (r = 0.55, P
4.Comparative study on cancer cell apoptosis between gastric and intestinal-type human gastric carcinoma.
Limin, LIU ; Shengjun, LU ; Qingshan, CHEN ; Heping, XIAO ; Yongrong, MAO
Journal of Huazhong University of Science and Technology (Medical Sciences) 2006;26(6):674-7
Apoptosis of cancer cells between the gastric and intestinal-type human gastric carcinoma were compared in terms of the expression of oncogene MDM2 and CD68, the histological types, the infiltration depth, and lymph node metastasis. Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling (TUNEL) assay was employed to stain apoptotic cells. Histochemical method(AB-PAS) was applied to stain mucus that is neutral or acidic in nature. Immunohistochemical method (SABC) was used to detect expression of MDM2 and CD6. The results showed that the mean apoptosis index (AI) of total 48 cases was 8.60+/-2.60. AI in the 30 intestinal type cases was significantly higher than that in the 18 gastric type cases (t=4.67, P<0.01). In the 30 intestinal type cases, the spontaneous apoptosis index of MDM2 negative cases was significantly higher than that of the positive cases (t=7.16, P<0.01). And in the 18 gastric type cases, the same result was found. (t=11.39, P<0.01). The MDM2 positive ratio in gastric type cases was higher than that in intestinal type cases (chi2=4.68, P<0.05). There is no significant difference in AI between cases of lymph node metastasis and non-metastasis cases in intestinal type cases (t=0.26, P>0.05). But in the gastric type cases, a significant difference existed (t=5.87, P<0.01). A significant difference in lymph node metastasis ratio was found between the two gastric carcinoma types (chi2=4.48, P<0.05). The CD68 expression ratio in the 30 intestinal type cases was much lower than that in the 18 gastric type cases (t=4.29, P<0.01). AI of 25 MDM2-positive cases was much lower than that of the 23 MDM2-negative cases (t=7.80, P<0.01). CD68 positive ratio in the 25 MDM2-negative cases was much lower than that in the 23 negative cases. The difference was statistically significant (t=10.90, P<0.01). Except for few cells scattering within the cancer nest, most CD68 positive cells infiltrated in the interstitium around the cancer tissue. In the high-AI cases, CD68-positive cells increased. And the CD68-positive cells decreased in low-AI cases (r=0.96, P<0.01). Logistic regression analysis suggested that among the control variables, only AI was a statistically significant factor in the regression model (chi2=9.64, P<0.01). We concluded that (1) the spontaneous apoptosis index in gastric-type cases of gastric carcinoma was significantly lower than that in intestinal type cases; (2) AI in the two types was influenced by the expression of MDM2 and lymph node metastasis, but no visible connection was found between AI and the infiltration depth or histological types; (3) in the intestinal type cases, AI and the CD68-positive cells increased in MDM2-negative cases.
5.Prevalence of chronic kidney disease and its related factors in chinese elderly persons
Biao LI ; Haimei QI ; Dongjie YU ; Yajing SHANG ; Defa CHU ; Limin MAO ; Pulin YU ; Hua WU
Chinese Journal of Geriatrics 2009;28(3):250-253
Objective To investigate the prevalence of chronic kidney disease (CKD) and its risk factors in Chinese elderly persons.Methods All of the people who underwent physical examination and treatment in the geriatric department of Beijing Hospital during January 2004 to January 2007 were included in the study.Age, body height, body mass index and blood pressure were recorded.Bloody urine was ascertained by phase-contrast microscope, and urine protein was measured by dipstick test.The hemoglobulin, serum eretinine, blood urea nitrogen, blood lipid and serum uric acid were measured by autobiochemical analyzer.HbsAg was checked by enzyme-linked immunosorbent assay (ELISA).Glomerular filtration rate (GFR) was estimated by Crockeroft-Gauh equation and abbreviated MDRD equation.Binary logistic regression analysis was used to test the risk factors for proteinuria and CKD.Results The prevalence of proteinuria was 4.9% in 1082 elderly persons.And 47.23% of the elderly suffered from decreased renal function.The morbidity of CKD was 48.43%.Binary logistic regression analysis showed that diabetes(OR= 2.257) and microscopic hematuria(OR=5.324) were the risk factors of proteinuria (both P<0.05).And the risk factors for CKD were hypertension(OR= 1.459), coronary arth'erosclerotic heart disease(OR=3.290), chronic obstructive lung diseases(OR=2.094), malignant tumor(OR=2.072), hyperuricemia(OR= 1.928),anemia(OR=8.122)and hematuria( OR= 1.604) (all P<0.05).Conclusions The morbidity of CKD in Chinese elderly persons was 48.43%.And the related risk factors were diabetes,hypertension, hyperuricemia, coronary artherosclerotic heart disease and chronic obstructive lung disease.
6.Preparation of a deactivation vaccine against Aeromonas hydrophila and research about its immue effect in the north of Henan
Jianyi GUAN ; Huili MAO ; Wenxu HE ; Limin YANG ; Yundong FU ; Jingmei ZHANG ; Jiajing WU ; Qianji NING
Chinese Journal of Immunology 2015;(11):1501-1504
Objective:Selected virulence factors more than and high virulent Aeromonas hydrophia strain made into inactivated vaccine,to study the immunization effect of inactivated vaccine.Methods:Crucian were vaccinated with formalin-killed vaccine via in-traperitoneal injection.Controls were injected with the same volumes of saline.Then the antibody titres, histopathology and relative percent survival were analyzed from samples of both groups.Results: The antibody in the indirect agglutination reaction could be detected in vaccinated fish once a weeks after immunization and reached highest level 6 weeks after immunization.The histopathology analysis indicated that the vaccine had a good protective effect on crucian target organs.Vaccinated fish showed 100%relative percent survival and the immune period would be 6 month.Conclusion:The vaccine in this study has a significant protective effect on crucian and may be used as effective fish vaccines against bacterial septicemia.
7.Biocompatibility of electrospun poly(lactide-co-glycolide)/polyethylene glycol nanofibrous scaffold with mouse neural stem cells
Chang LIU ; Limin RONG ; Shangfu LI ; Mao PANG ; Yang YANG ; Bin LIU
Chinese Journal of Tissue Engineering Research 2014;(47):7585-7590
BACKGROUND:Poly(lactide-co-glycolide) (PLGA) scaffold is widely used in tissue engineering, but its poor cel adhesion ability and strong hydrophobicity limit its further development and application. OBJECTIVE: To study the biocompatibility of electrospun poly (lactide-co-glycolide)/polyethylene glycol (PLGA-PEG) nanofibrous scaffolds with mouse neural stem celsin vitro. METHODS:Neural stem cels were isolated from embryos of CD-1 mice at 15 embryonic days. Electrospinning was used to prepare PLGA and PLGA-PEG nanofibrous scaffolds. Scanning electron microscope was used for scanning observation of scaffolds. The 5th passage neural stem cels were seeded onto PLGA and PLGA-PEG scaffolds respectively, and culturedin vitro. RESULTS AND CONCLUSION: Interconnected porous network structure was observed in both two kinds of scaffolds under the scanning electron microscope. Fiber diameters and porosities of PLGA and PLGA-PEG scaffolds showed no significant differences (P > 0.05). Cel Counting Kit-8 detection showed neural stem cels grew wel on both two kinds of scaffolds and the absorbance value of two groups increased continuously with incubation time (1, 3, 5, 7, 9, 11 days). And there were statisticaly significant differences in the absorbance values between two groups at each time point (P < 0.05). Moreover, the cel adhesion rate was significantly higher in the PLGA-PEG group than in the PLGA group at 3, 6, 9 hours of culture (P < 0.05). Hoechst 33342 staining showed normal morphology and quality of the nuclei, and significantly more cels were observed in the PLGA-PEG group than the PLGA group (P < 0.05). Under the scanning electron microscope, compared with the PLGA scaffold, the PLGA-PEG scaffold was better for growth and matrix secretion of neural stem cels. In conclusion, PLGA-PEG nanofibrous scaffolds prepared by electrospinning are safe, non-toxic and suitable for neural stem cels growth with wel biocompatibility, appropriate aperture and porosity.
8.Nine patients with paratyphoid fever A complicated with severe kidney damage
Limin CAI ; Minghe YAN ; Xitian HUANG ; Wenzhong MAO ; Yunsheng LI ; Xi LIN
Chinese Journal of Emergency Medicine 2006;0(01):-
Objective To study the clinical characteristics,diagnosis,and treatment of paratyphoid fever A (fever A) complicated with severe kidney damage. Methods The data were retrospectively reviewed.Results The symptoms of poisoning and gastrointestinal symptoms as well as the slightly abnormal urinalysis (proteinuria, hematuria) appeared early, which got worse along with the progression of the disease. The symptoms of urinary system and azotemia appeared in the worst period. After treatment with antibiotic, 4 patients recovered early.Conclusion Paratyphoid fever A can cause severe kidney damage, with non-specific symptoms at the early stage. The seriousness of the abnormal of urine test can't be ignored as those provides the basis for the early diagnosis. The early use of antibiotic is the key to avoid and lessen the serverity of kidney damage.
9.Association of serum uric acid level with the prevalence of metabolic syndrome
Yanjia XU ; Jing LIU ; Qi ZHANG ; Mao LI ; Limin TIAN ; Jinxing QUAN
The Journal of Practical Medicine 2016;32(15):2556-2559
Objective To investigate the correlation of serum uric acid (SUA) level with the prevalence of metabolic syndrome (MS). Methods A random, multi-stage cross-sectional study was performed in 2364 resuden, aged from 20 to 74, in Lanzhou, Gansu province.The resudents were divided into groups according to the level of SUA or 4 components of MS. The relationship between SUA and MS was analyzed. Results The overall morbidity of MS was 20.13 % in Lanzhou , and the morbidity of MS was higher in female than that in male (23.22% vs. 17.37%, P < 0.05);The prevalences of MS was higher in the hyperuricemia (HUA) people than that in the healthy people (32.40 % vs. 17.70 %,P < 0.05); With the increase of SUA level, the morbidity rates of MS and its components increased;The risk of MS showed a gradual increase with the SUA level, the risk of MS(odds ratio,95%CI)in sex-specific quartiles of SUA was 2.33(1.45 ~ 3.71),2.33(1.45 ~3.71),2.44(1.53 ~ 3.89)in male;and 1.58(0.97~2.56),2.54(1.60 ~ 4.00),5.29(3.41 ~ 8.22)in female. The more the components of MS , the higher the SUA level was found for both the male and the female subjects , with the higher rate of HUA morbidity(P < 0.05). Conclusion There is a significant association between SUA level and MS, as well as the MS components. Comprehensive prevention and control should be taken for the reduction of the risk factors, and much attention should be paid to monitor the SUA level.
10.Comparative Study on Cancer Cell Apoptosis between Gastric and Intestinal-type Human Gastric Carcinoma
Limin LIU ; Shengjun LU ; Qingshan CHEN ; Heping XIAO ; Yongrong MAO
Journal of Huazhong University of Science and Technology (Medical Sciences) 2006;26(6):674-677
Apoptosis of cancer cells between the gastric and intestinal-type human gastric carcinoma were compared in terms of the expression of oncogene MDM2 and CD68, the histological types, the infiltration depth, and lymph node metastasis. Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling (TUNEL) assay was employed to stain apoptotic cells.Histochemical method(AB-PAS) was applied to stain mucus that is neutral or acidic in nature. Immunohistochemical method (SABC) was used to detect expression of MDM2 and CD6. The results showed that the mean apoptosis index (AI) of total 48 cases was 8.60±2.60. AI in the 30 intestinal type cases was significantly higher than that in the 18 gastric type cases (t=4.67, P<0.01). In the 30intestinal type cases, the spontaneous apoptosis index of MDM2 negative cases was significantly higher than that of the positive cases (t=7.16, P<0.01). And in the 18 gastric type cases, the same result was found. (t=11.39, P<0.01). The MDM2 positive ratio in gastric type cases was higher than that in intestinal type cases (x2=4.68, P<0.05). There is no significant difference in AI between cases of lymph node metastasis and non-metastasis cases in intestinal type cases (t=0.26, P>0.05). But in the gastric type cases, a significant difference existed (t=5.87, P<0.01). A significant difference in lymph node metastasis ratio was found between the two gastric carcinoma types (x2=4.48, P<0.05).The CD68 expression ratio in the 30 intestinal type cases was much lower than that in the 18 gastric type cases (t=4.29, P<0.01). AI of 25 MDM2-positive cases was much lower than that of the 23MDM2-negative cases (t=7.80, P<0.01). CD68 positive ratio in the 25 MDM2-negative cases was much lower than that in the 23 negative cases. The difference was statistically significant (t=10.90,P<0.01). Except for few cells scattering within the cancer nest, most CD68 positive cells infiltrated in the interstitium around the cancer tissue. In the high-AI cases, CD68-positive cells increased. And the CD68-positive cells decreased in low-AI cases (r=0.96, P<0.01). Logistic regression analysis suggested that among the control variables, only AI was a statistically significant factor in the regression model (x2=9.64, P<0.01). We concluded that (1) the spontaneous apoptosis index in gastric-type cases of gastric carcinoma was significantly lower than that in intestinal type cases; (2) AI in the two types was influenced by the expression of MDM2 and lymph node metastasis, but no visible connection was found between AI and the infiltration depth or histological types; (3) in the intestinal type cases,AI and the CD68-positive cells increased in MDM2-negative cases.