1.Activation of JAK-STAT1 signal transduction pathway in lesional skin and monocytes from patients with systemic lupus erythematosus
Jie LI ; Shuang ZHAO ; Mei YI ; Xing HU ; Ji LI ; Hongfu XIE ; Wu ZHU ; Mingliang CHEN
Journal of Central South University(Medical Sciences) 2011;36(2):109-115
Objective To study the activation of Janus protein tyrosine kinase (JAK)/signal transducer and activator of transcription 1 (STAT1) signaling pathway and its inhibitor-signal transducer and activator of transcription-1(SOCS-1) in patients with systemic lupus erythematosus. Methods A total of 45 patients with active systemic lupus erythematosus (SLE) and 30 healthy controls were randomly selected. Western blot was performed to measure the expression of Stat1 protein and phospho-Stat1 protein (an activated form of Stat1 protein) in the monocytes after stimulation with recombinant high mobility group box1 (rHMGB1) at various time points. Expression of Stat1 protein in the skin or lesional skin was also detected. Phasic expressions of SOCS-1 mRNA in the monocytes after rHMGB1 stimulation were detected by real-time reverse transcription-polymerase chain reaction. SOCS-1 gene expression in the skin or lesional skin was also detected. Results The expression level of Stat1 proteins in the monocytes from patients with SLE was higher than that from healthy controls (t=9.16,P<0.01) and positively correlated with SLE disease activity index (SLEDAI) (r=0.59,P<0.01). Expression of phospho-Stat1 in the monocytes from SLE patients was time-dependently upregulated after stimulation with rHMGB1 at various time points, while expression of SOCS-1 mRNA remained unchanged(all P>0.05). Expressions of phospho-Stat1 protein and SOCS-1 mRNA in the monocytes from healthy controls were increased transiently after stimulation with rHMGB1(all P<0.05). Both expressions of phospho-Stat1 protein and SOCS-1 gene in the lesional skin from patients with SLE were upregulated compared with those in normal skin from healthy controls (all P<0.01). Conclusion There are hyperactivation of JAK-STAT1 signaling pathway and negative feedback down-regulation of SOCS-1 in patients with systemic lupus erythematosus. HMGB-1 may be partly involved in the pathogenesis of SLE by the abnormal mediating function of JAK-STAT1 signal transduction pathway.
2.Analysis of nutritional status of iodine among residents in Chongqing in 2009
Shuang, ZHOU ; Bang-zhong, XIAO ; Cheng-guo, WU ; Xing-jian, LUO ; Ya-lin, CHEN ; Xin-shu, LI ; Jing, CHEN
Chinese Journal of Endemiology 2011;30(3):312-315
Objective To investigate the nutritional status of iodine among residents in Chongqing, and to facilitate scientific prevention and control of iodine deficiency disorders. Methods Select 9 towns in each of the 40 districts (counties) in Chongqing, and collect 40 resident edible salt samples in each of the selected town to detect salt iodine by direct titrimetry. Select 5 towns on the site of the east, west, south, north and middle of every district (county), select 20 children aged 8 to 10 in each of the selected town to collect urine samples and detect urinary iodine by As-Ce catalytic spectrophotometric assay. Results The median of iodine of 14 217 salt specimens by household was 292 mg/kg with a coverage rate of qualified iodized salt of 98.90%( 14 061/14 217). The consumption rate of qualified iodized salt was 95.59%( 13 590/14 217). The median of urinary iodine for 4050 children aged 8 to 10 was 247.20μg/L, of which < 50 μg/L accounted for 4.60%(186/4050), 50-99μg/L accounted for 7.28% (295/4050), 100 - 199 μg/L accounted for 26.44% (1071/4050), 200 - 299 μg/L accounted for 25.58% (1036/4050), 300 μg/L or more, accounted for 36.10% (1462/4050). However, no significant difference was observed between different age groups(x2 = 3.77, P > 0.05). At district (county) level, the median of urinary iodine in 10(25.00%) districts (counties) was 100 - 200 μg/L, that in other 23(57.50%) districts (counties) was 200 - 300 μg/L, and that in other 7(17.50%) districts/counties was greater than 300 μg/L, and statistical significance was observed between different districts/counties (x2 = 441.95, P < 0.01). Conclusions Current iodine nutrition among residents in Chongqing is adequate. While there is excess, need to reduce the amount of salt iodization.
3.Association between methylthioadenosine phosphorylase gene single nucleotide polymorphisms and myocardial infarction in Chinese Han ethnicity
Qi ZHANG ; Shuang-Shuang CHENG ; Jie YAN ; Xing-Dong CHEN ; Xiao-Feng WANG ; Xin-Chun YANG
Chinese Journal of Epidemiology 2010;31(1):83-86
Objective To investigate the association between methylthioadenosine phosphorylase (MTAP) gene single nucleotide polymorphisms (SNP) and myocardial infarction (MI) in the Chinese Han ethnicity. Methods 432 patients suffered from myocardial infarction and 430 controls were involved for case and control groups, respectively. Nine tag SNPs in MTAP gene were selected and genotyped. Results We found no significant association of selected tag SNPs with MI in all of the samples. However, in stratified analysis, significant association was observed at rs7027989 in male subjects. The risk of MI increased by 26% (P=0.005) for male subjects of minor allele carriers in a dominant model. The increased risk of MI at rs7027989 remained significant after adjusting for confounding factors. Conclusion MTAP gene might be involved in the etiology of MI in Chinese Han ethnicity.
4.The study of catheterization to measure ventricular pressure in mice through venous cannula in external jugular vein.
Pan-Pan LIU ; Shuang-Quan YAN ; Ma-Yun CHEN ; Li-Zhen ZOU ; Xiao-Ying HUANG ; Liang-Xing WANG
Chinese Journal of Applied Physiology 2014;30(2):189-192
OBJECTIVETo study a feasible method of measuring right ventricular pressure by catheterization in mice.
METHODSMeasuring the right ventricular pressure and the pulmonary artery pressure by homemade PE pipe through venous cannula in external jugular vein, using catheterization in mice with powerlab multimodal biometric signal recording system.
RESULTSForty-six out of 51 mice were experimented with this method smoothly and got a total success rate of 90.2%. Thirty of 33 normal mice and 16 of 18 mice with pulmonary arterial hypertension (PAH) were catheterized successfully. The right ventricular pressure were as follow: systolic blood pressure: (23.4 +/- 5.7) mmHg in normal group vs (32.2 +/- 2.8) mmHg in mice with PAH, diastolic blood pressure: (3.7 +/- 2.6) mmHg vs (3.8 +/- 2.0) mmHg, mean pressure: (12.0 +/- 3.7) mmHg vs (14.9 +/- 2.3) mmHg. After autopsy for those 5 failed cases, we found that 2 cases were into the inferior vena cava, another 2 cases pierced the right auricle and the last one punctured the axillary vein into the chest wall.
CONCLUSIONMeasuring the right ventricular pressure through venous cannula in external jugular vein with homemade PE pipe in mice gets not only a high success rate but also help to save time. Moreover, this method can be popularized easily. It is a good and feasible method for measuring right ventricular pressure in mice.
Animals ; Cardiac Catheterization ; methods ; Jugular Veins ; Male ; Mice ; Mice, Inbred BALB C ; Ventricular Pressure
5.Mechanism of sophocarpine in treating experimental colitis in mice.
Jian-mei ZHANG ; Ya-bi ZHU ; Xing DENG ; Chang-xiong WANG ; Shuang-mei LUAN ; Yue-xiang CHEN
China Journal of Chinese Materia Medica 2015;40(15):3081-3087
To study the preventive effect of sophocarpine (Soc) on dextran sulfate sodium (DSS)-induced colitis in mice, in order to analyze the influence of Soc on toll like receptor 4 (TLR4)/mitogen-activated protein kinases (MAPKs) and janus tyrosine kinase 2 signal transducer and activator of transcription 3 (JAK2/STAT3) signal pathways in mice intestinal tissues. The mice was given 2.5% DSS for 6 days to induce the acute colitis model. The Soc-treated group was intraperitoneally injected with sophocarpine 30 mg · kg(-1) · d(-1) since the day before the experiment to the end. The disease activity index (DAI) was assessed everyday, and the colonic morphology and histological damage were observed with HE staining. The mRNA expressions of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6) were detected by real-time RT-PCR. The changes in key protein kinase p38 mitogen-activated protein kinase (p38MAPK), c-Jun NH2-terminal protein kinase1/2 (JNK1/2), extracellular signal-regulated kinase1/2 (ERK1/2), JAK2, STAT3 in TLR4/MAPKs and JAK2/STAT3 signaling pathways were detected by western blot. The result showed that the model group showed statistical significance in body weight, DAI, colon length and histopathological changes compared with the normal group (P <0.05); however, the Soc-treated group showed significant improvements in the above indexes compared with the model group (P <0.05). TNF-α, IL-1β and IL-6 in the model group was significantly higher than that in the normal group (P <0.05), but lowered in the Soc-treated group to varying degrees (P <0.05). In the normal group, the expressions of TLR4 and the phosphorylation of P38, JNK1/2, JAK2, STAT3 were at low levels; in the model group, the phosphorylation of P38, JNK1/2, JAK2, STAT3 increased; the Soc-treated group showed a decrease in TLR4 expression compared with the model group, with notable declines in the phosphorylation of TLR4, P38, JNK1/2, JAK2, STAT3. These findings indicate that Soc can inhibit TLR4/MAPKs, K2/STAT3 signaling pathway activation, reduce the expression of proinflammatory cytokines TNF-α, IL-1β and IL-6 and relieve inflammatory reactions, so as to effectively prevent experimental colitis.
Alkaloids
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pharmacology
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therapeutic use
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Animals
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Colitis
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drug therapy
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immunology
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pathology
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Cytokines
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genetics
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Janus Kinase 2
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antagonists & inhibitors
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physiology
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Male
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Mice
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Mice, Inbred BALB C
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Phosphorylation
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STAT3 Transcription Factor
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antagonists & inhibitors
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physiology
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Toll-Like Receptor 4
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antagonists & inhibitors
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physiology
6.The early change of hematopoietic injury induced by high dose 60Co γ-rayirradiation in rhesus
Ruiying ZHANG ; Hongling OU ; Guolin XIONG ; Shuang XING ; Suozhu SUN ; Shanshan DING ; Fenghua CHEN ; Zuying YU ; Xinru WANG
International Journal of Laboratory Medicine 2017;38(15):2022-2024,2027
Objective To observe the early changes of related indexes after high dose of 60Co γ-ray irradiation on rhesus monkey hematopoietic system.Methods A total of 33 rhesus monkeys were randomly divided into normal control and different irradiation control group,rhesus monkeys in irradiation control group were given different doses(4,8,12 Gy) irradiation to establish acute radiation sickness(ARS) models.XE-2100 automatic blood cell analyzer detected the peripheral blood before and after the irradiation of 3,6,9,12,24,48,80 h.The rhesus monkeys were sacrificed to have a observation of sternum pathological changes at 6,48 and 80 h after 4,8,12 Gy 60Co γ-ray irradiation.Results The number of white blood cell in peripheral blood of the rhesus monkeys after 4 and 8 Gy 60Co γ-ray irradiation were lower than that before irradiation at 3 h after irradiation,as was significant increased at 6 h after irradiation,the highest values were 136.04%.and 221.38% after 9 h(with before irradiation values was 100.00%,the same below),become obviously drooped from 12 h after irradiation,show clearly temporary peak.But the number of white blood cell after 12 Gy 60Co γ-ray irradiation was significant increased at 6 h after irradiation,at the highest of 9 h,become obviously drooped from 12 h after irradiation.Peripheral blood neutrophile count was significant increased at 6 h after irradiation,at the highest of 9 h,become obviously drooped from 12 h after irradiation.Peripheral blood lymphocyte count fell sharply after irradiation,3 h detection value was only 12.02%-25.04% of before irradiation.Sternal bone marrow nucleated cell number decreased sharply after irradiation,the more irradiation dose,the less residual hematopoietic cells.Conclusion In the early stage of BM-ARS,temporary peaktime node of the white blood cell and neutrophil count could be regarded as the best delivery time of hematopoietic cytokine therapy.
7.Repair of damaged intestinal mucosa in a mouse model of sepsis
Rui-Ming CHANG ; Li-Qiang WEN ; Jian-Xing CHANG ; Yu-Ru FU ; Zhi-Peng JIANG ; Shuang CHEN
World Journal of Emergency Medicine 2013;4(3):223-228
BACKGROUND:The intestine is not only the main target attacked by sepsis but also the vital organ which mediated sepsis. The recovery of the damaged intestinal barrier structure and function is related to the occurrence and outcome of multiple organ dysfunction syndrome (MODS). How to protect and reduce the damage of the intestinal mucosa and how to promote the reconstruction of the intestinal mucosa have been the important topics in sepsis for many years. This study aimed to investigate the influential factors of intestinal mucosal reconstruction after intestinal epithelial injuryin vivo in a mouse model of sepsis.METHODS:Mice were subjected to cecal ligation and puncture (CLP) for induction of sepsis to assess intestinal mucosal damage, epithelial cell apoptosis, and transformed number of goblet cells, and to detect the concentration of TNF-α, IL-1 and TGF-β1 and TFF3 (trefoil factor 3) expression in the small intestinal mucosa. All above were performed by HE staining, western blot, ELISA and immunohistochemistry respectively. The experimental animals were divided into a sepsis group and a sham-operation group. The animals with sepsis were separately killed at 6 (7 animals), 24 (7 animals) and 48 hours (7 animals) after CLP.RESULTS:Injured intestinal mucosa was observed in the 3 groups under a light microscope, in which damage scores in the 24-hour and 48-hour groups were higher than in the 6-hour group and no difference was found between the two groups. Moreover, less of goblet cells or other epithelial cells adjacent to the injured surface migrated into the wound to cover the denuded area. The number of goblet cells was substantially decreased in the three CLP groups compared with the sham-operation group. Protein levels of IL-1 and TNF-α were significantly increased by 3-4 fold at all time points when compared with the sham-operation group, and cleaved caspase-3 by 4 fold. Although TFF3 expression was modestly increased for 6 hours after the onset of CLP, it appeared to decline at 24 hours and 48 hours as shown by Western blot. A similar tendency was observed upon TGF-β1, i.e. the protein level was not elevated at 24 hours and 48 hours, but increased modestly at 6 hours.CONCLUSIONS:Sepsis from CLP shows less restitution on the surface of injured intestinal mucosa. There is evidence that both constant inflammatory reaction and epithelial cell apoptosis may affect mucosal reestablishment of the intestine at the onset of sepsis. Mucosa after severe sepsis showed the state of high inflammation, and declined goblet cell function and mucosal reconstruction, which affected the repair of damaged intestinal barrier. Constant inflammatory reaction, and declined goblet cell function and mucosal reconstruction ability may affect the reestablishment of intestinal mucosa at the onset of sepsis.
8.The water-soluble TF3 component from Eupolyphaga sinensis Walker promotes tibial fracture healing in rats by promoting osteoblast proliferation and angiogenesis
Binghao Shao ; Xing Chen ; Jin' ; ge Du ; Shuang Zou ; Zhaolong Chen ; Jing Wang ; Huaying Jiang ; Ruifang Lu ; Wenlan Wang ; Chunmei Wang
Journal of Traditional Chinese Medical Sciences 2024;11(2):245-254
Objective:
To determine the active components of Eupolyphaga sinensis Walker (Tu Bie Chong) and explore the mechanisms underlying its fracture-healing ability.
Methods:
A modified Einhorn method was used to develop a rat tibial fracture model. Progression of bone healing was assessed using radiological methods. Safranin O/fast green and CD31 immunohistochemical staining were performed to evaluate the growth of bone cells and angiogenesis at the fracture site. Methylthiazoletetrazolium blue and wound healing assays were used to analyze cell viability and migration. The Transwell assay was used to explore the invasion capacity of the cells. Tubule formation assays were used to assess the angiogenesis capacity of human vascular endothelial cells (HUVECs). qRT-PCR was used to evaluate the changes in gene transcription levels.
Results:
Tu Bie Chong fraction 3 (TF3) significantly shortened the fracture healing time in model rats. X-ray results showed that on day 14, fracture healing in the TF3 treatment group was significantly better than that in the control group (P = .0086). Tissue staining showed that cartilage growth and the number of H-shaped blood vessels at the fracture site of the TF3 treatment group were better than those of the control group. In vitro, TF3 significantly promoted the proliferation and wound healing of MC3T3-E1s and HUVECs (all P < .01). Transwell assays showed that TF3 promoted the migration of HUVECs, but inhibited the migration of MC3T3-E1 cells. Tubule formation experiments confirmed that TF3 markedly promoted the ability of vascular endothelial cells to form microtubules. Gene expression analysis revealed that TF3 significantly promoted the expression of VEGFA, SPOCD1, NGF, and NGFR in HUVECs. In MC3T3-E1 cells, the transcript levels of RUNX2 and COL2A1 were significantly elevated following TF3 treatment.
Conclusion
TF3 promotes fracture healing by promoting bone regeneration associated with the RUNX2 pathway and angiogenesis associated with the VEGFA pathway.
9.Effect of (S)-4C3HPG on brain damage in the acute stage of moderate traumatic brain injury model of mice and underlying mechanism.
Nan YANG ; Shuang-Shuang DAI ; Ya-Lei NING ; Xing-Yun CHEN ; Yan ZHAO ; Ping LI ; Yuan-Guo ZHOU
Acta Physiologica Sinica 2010;62(6):555-559
The aim of this study is to investigate the effect of (S)-4-carboxy-3-hydroxy-phenylglycine [(S)-4C3HPG], a mixed group I glutamate metabotropic receptor antagonist and a group II agonist, on impairment in a cortical impact model of traumatic brain injury (TBI) in mice and to elucidate the possible mechanisms. Mice were injected (i.p.) with saline, 1 mg/kg (S)-4C3HPG, 5 mg/kg (S)-4C3HPG and 10 mg/kg (S)-4C3HPG (n=10 per group), respectively, at 30 min before moderate TBI. Neurological deficit scores, water content in injured brain and glutamate concentration in cerebral spinal fluid (CSF) were detected at 24 h after TBI. The expressions of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) mRNA in injured cortex were also detected by real-time RT-PCR. The results showed that the neurological deficits and cerebral edema were significantly attenuated in mice pretreated with (S)-4C3HPG (5 and 10 mg/kg respectively) compared with those in mice pretreated with saline. Furthermore, (S)-4C3HPG treatment also decreased the glutamate concentration in CSF and the expressions of TNF-α and IL-1β mRNA remarkably in a dose-dependent manner. These results suggest that (S)-4C3HPG treatment attenuates cortical impact-induced brain injury possibly via suppression of glutamate release and inhibition of excessive inflammatory cytokine production. These findings highlight the potential benefit of glutamate metabotropic receptor ligand for preventing TBI.
Animals
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Brain Injuries
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drug therapy
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metabolism
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physiopathology
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Cytokines
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metabolism
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Glutamic Acid
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cerebrospinal fluid
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Glycine
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analogs & derivatives
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therapeutic use
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Male
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Mice
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Mice, Inbred C57BL
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Receptors, Metabotropic Glutamate
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agonists
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antagonists & inhibitors
10.Expression and its clinical significance of RBP4 in vitreous in patients with proliferative diabetic retinopathy
Tian-Yue CAO ; Ming AI ; Hong-Xia YANG ; Zhen CHEN ; Yi-Qiao XING ; Shuang-Hong JIANG
International Eye Science 2018;18(5):811-814
AIM:To observe the concentration of RBP4 and IL-6 in vitreous of proliferative diabetic retinopathy (PDR). METHODS: A total of 65 patients (66 eyes) were enrolled in Department of Ophthalmology, Renmin Hospital of Wuhan University from February 2017 to July 2017 with the informed consent. The patients were divided into PDR group (23 cases) and NPDR group (16 cases). Twenty- six patients without diabetic mellitus (DM) served as control group. The demography was matched among the groups, but the course of DM, the blood glucose level and the HbA1c level were elevated in the PDR group and the NPDR group (all P<0.05). Vitreous samples were collected during the procedure of vitrectomy. RBP4,IL-6,TNF-α concentrations in vitreous specimens were detected by ELISA. The differences of vitreous RBP4, IL-6 and TNF-α in various groups were statistically analyzed by ANOVA, respectively. The correlations between RBP4 and IL - 6, TNF - α were calculated by Pearson correlation analysis. RESULTS:The concentration of RBP4 in PDR group,the NPDR group and control group were 13.68士2.66, 11 03士1 12,10.45士1.17μ g/Ml, and the concentration of IL-6 were 56.0士10.27, 20.92士5.77, 10.26士1.91pg/Ml. RBP4 and IL- 6 concentrations were elevated in PDR group compared with NPDR group and control group, with significant difference among three groups (F = 12. 135, 161.167; P < 0. 01). IL - 6 concentrations in vitreous increased in the NPDR group in comparison with control group(P<0.05). RBP4 concentrations had no significant difference between the NPDR group and the group(P>0 05). Pearson correlation coefficient was significant positive between RBP4 concentration and IL - 6 concentration(r=0.606,P=0.001). CONCLUSION: RBP4 is probability involved in the inflammation pathogenesis of PDR. These results indicate that RBP4 could be a new target for the diagnosis and treatment of PDR.