Background: Zhibai Dihuang Granule (ZDG) is a traditional Chinese medicine which has been used to treat Yin-deficiency- heat (YDH) syndrome for thousands of years in China. However, little work has been conducted to explore the molecular mechanism of ZDG in YDH syndrome, and the processes of YDH syndrome prevention and treatment have been developed slowly. The present study was aimed to explore the therapeutic mechanism of ZDG on YDH syndrome. Methods: The YDH syndrome rats were induced by hot Chinese herbs, then treated by ZDG orally for 1 week. Body weight was measured every 2 days. After sacrifice, blood samples were collected and the thymus, adrenal glands, spleen, and liver were immediately removed and weighed. iTRAQ-based proteomics approach was applied to explore the serum protein alterations with the treatment of ZDG, and to investigate the underlying mechanism of ZDG in treating YDH syndrome. Results: The body weights of YDH syndrome rats were significantly decreased compared with control group, and increased in ZDG treated rats. The relative weights of thymus in YDH syndrome rats were increased compared with the control rats, and significantly decreased in after ZDG treatment. In the proteomic analyses, seventy-one proteins were differentially expressed in the YDH syndrome group and the ZDG treated group, including 10 up-regulated and 61 down-regulated proteins. Gene ontology analysis revealed that the differentially expressed proteins were mostly related to immune response, and pathway enrichment analysis showed that these proteins were enriched in coagulation and complement cascades. Enzyme-linked immunosorbent assay was performed to detect the protein levels in coagulation and complement cascades, and the results showed that complement component 5 levels were significantly increased, while fibrinogen gamma chain levels were significantly decreased in the ZDG treated group. Conclusions: We found that ZDG treatment could lead to proteins alteration in immune response, especially in coagulation and complement cascades. ZDG can up-regulate the proteins in the complement cascade to eliminate pathogens, and down-regulate the proteins in the coagulation cascade to suppress inflammation. Our study provides experimental basis to understand the therapeutic mechanism of ZDG and revealed that ZDG can regulate coagulation and complement cascades in treating YDH syndrome.

Yin and Yang are the two counter‐balancing aspects in ancient Chinese philosophy. In traditional Chinese medicine, Yin deficiency syndrome (YDS) is a common sub‐health state with complex causes. While the syndrome may be treated to various degrees of effectiveness with traditional Chinese medicine, efficient modern methods are yet to be developed for diagnosing and treating the YDS. Here we performed a metabolomics study on YDS in rats. Serum metabolites in rats were analyzed using ultra‐performance liquid chromatography‐mass spectrometry (UPLC–MS) method to identify potential biomarkers for YDS. The rats were divided randomly into the healthy control group, the untreated YDS group, and the anemarrhena treated YDS group. Compared with the control group, significant increase in the metabolites such as dihydrotestosterone (DHT) and 5β‐DHT, 4‐imidazolone‐5‐propanoate, 4‐(L‐alanin‐3‐yl)‐2‐hydroxy‐cis,cis‐muconate 6‐semialdehyde, and 5‐(L‐alanin‐3‐yl)‐2‐hydroxy‐cis,cis‐muconate 6‐semialdehyde were observed in the serum of untreated YDS group, which returned to normal in the anemarrhena treated group. Therefore, these metabolites may serve as potential biomarkers for YDS, and may facilitate the diagnosis and treatment of YDS.

Ethnopharmacological relevance: Zhibai Dihuang Granule (ZDG), a traditional Chinese medicine (TCM) made from eight Chinese herbs, has been classically used to treat Yin-deficiency-heat (YDH) syndrome. ZDG is well known with the therapeutic efficacy of nourishing Yin and decreasing internal heat in clinic, but the mechanism of ZDG's therapeutic effect is still not clear. Materials and methods: High doses of triiodothyronine (T3) were given intraperitoneally to induce Hyperthyroid YDH syndrome in SD rats. The animals were then treated with ZDG for one week. The iTRAQ-coupled with two-dimensional liquid chromatography-tandem mass spectrometry (2D LC-MS/MS) technique was used to screen the differentially expressed serum proteins between ZDG treated rats and YDH syndrome rats. The differentially expressed proteins were analyzed by bioinformatics method and were verified by enzyme-linked immunosorbent assay (ELISA). Results: A total of 55 differentially expressed proteins were identified, including 23 up-regulated proteins (> 1.25 fold, p < 0.05) and 32 down-regulated proteins (< 0.80 fold, p < 0.05). Among the differentially expressed proteins, 26 proteins returned to normal after ZDG treatment. Bioinformatics analysis showed that these proteins were mainly involved in immune response, including regulation of immune system process, complement activation, and humoral immune response mediated by circulating immunoglobulin. ELISA revealed significantly increased levels of Zinc-alpha-2-glycoprotein (Azgpl), L-selectin, C-reactive protein (Crp), Plasminogen (Pig), Kininogen 1 (Kngl), and significantly decreased levels of Mannose binding lectin 2 (Mb12) and Complement Clqb chain (Clqb) in ZDG treated rats compared with YDH syndrome rats. Bioinformatics analyses indicated that Azgpl participated in antigen processing and presentation, Crp, Clqb, and Mb12 were involved in complement activation, while L-selectin, Plg, and Kngl were involved in regulating the inflammatory response. Conclusions: Our study provides experimental evidence to understand the therapeutic mechanism of ZDG in YDH syndrome. The results suggested that ZDG may regulate the complement activation and inflammatory response, and promote the ability to recognize antigens to alleviate YDH syndrome.

Safranal, a main component of Crocus sativus, is suggested to have neuroprotective effects. The aim of this study was to investigate the effect of safranal and nanostructured lipid vehicle (NLV) carried safranal in acute and chronic experimental mice models of epilepsy. In PILO acute seizure model, safranal dose-dependently extended latency to generalized seizure, decreased the highest seizure stages and the number of generalized seizures. Moreover, NLV carried safranal further enhanced the anti-seizure effect, which is comparable to the action of sodium valproate. Meanwhile, NLV carried safranal reduced and delayed the electroencephalogram spectra power after pilocarpine injection. In histological aspect, safranal dose-dependently reduced the loss of neurons induced by seizure and NLV system further improved this protection at the same dose. In MES acute model, safranal markedly increased the electroconvulsive threshold, where NLV further improved its effect. In PTZ chronic seizure model, NLV carried safranal significantly delayed the kindling rate of progress and the time it took to reach generalized seizures as compared to NLV control group. In conclusion, this study indicates that safranal inhibits generalized seizure in acute and chronic epilepsy models in mice and NLV can enhance this effect. So, NLV carried safranal may have potential value in treatment of generalized epilepsy.

Rapid and efficient methods for the determination of cured tuberculosis (TB) are lacking. A total of 85 differentially expressed serum proteins were identified by iTRAQ labeling coupled with two-dimensional liquid chromatography-tandem mass spectrometry (2D LC-MS/MS) analysis (fold change >1.50 or <0.60, P < 0.05). We validated albumin (ALB), Rho GDP-dissociation inhibitor 2 (ARHGDIB), complement 3 (C3), ficolin-2 (FCN2), and apolipoprotein (a) (LPA) using the enzyme-linked immunosorbent assay (ELISA) method. Significantly increased ALB and LPA levels (P = 0.036 and P = 0.012, respectively) and significantly reduced ARHGDIB, C3, and FCN2 levels (P < 0.001, P = 0.035, and P = 0.018, respectively) were observed in cured TB patients compared with untreated TB patients. In addition, changes in ALB and FCN2 levels occurred after 2 months of treatment (P < 0.001 and P = 0.030, respectively). We established a cured TB model with 87.10% sensitivity, 79.49% specificity, and an area under the curve (AUC) of 0.876. The results indicated that ALB, ARHGDIB, C3, FCN2, and LPA levels might serve as potential biomarkers for cured TB. Our study provides experimental data for establishing objective indicators of cured TB and also proposes potential markers for evaluating the efficacy of anti-TB drugs.

本文主要探究在联合型恐惧学习记忆过程中激活小鼠运动皮层对学习记忆的影响.光遗传激活运动皮层后,比较转基因小鼠和同窝阴性小鼠的短时记忆,发现转基因小鼠在有声时僵直比率显著性高于同窝阴性小鼠;在训练环境中的僵直比率没有明显差异.在长期记忆中,转基因小鼠在有声时僵直比率显著性高于同窝阴性小鼠;在训练环境中的僵直比率没有明显差异.实验数据说明激活运动皮层会损伤小鼠的联合型学习记忆,而对小鼠的环境学习记忆没有影响.

Zhibai Dihuang Granule (ZDG) is a classical traditional Chinese medicine that has been used to treat yin‐deficiency‐heat (YDH) syndrome. However, the therapeutic mechanism of ZDG is still unclear. The aim of this study was to observe the therapeutic effect and mechanism of ZDG in hyperthyroidism yin‐deficiency rats by detecting the serum proteome expression. Triiodothyronine (T3) was used to induce the hyperthyroidism yin‐deficiency model of rats, and the rats were treated daily with ZDG by gastrogavage. In the proteomics experiment, a total of 93 differentially expressed serum proteins (49 up‐regulated proteins and 44 down‐regulated proteins) were identified by iTRAQ labeling coupled with two‐dimensional liquid chromatography‐tandem mass spectrometry (2D LC‐MS/MS) analysis (fold change >1.50 or <0.60, p < 0.05) between model rats and control rats. After ZDG treatment, 16 proteins among up‐regulated proteins showed obvious downward trend, and 10 proteins among down‐regulated proteins showed obvious upward trend. Bioinformatics analysis of the proteins which returned to normal level revealed that 10 of these proteins were involved in immune response, especially in complement and coagulation cascades. After ZDG treatment, L‐selectin, C‐reactive protein (CRP), and Zinc‐alpha‐2‐glycoprotein (Azgp1) levels were significantly increased (p=0.0002, p=0.0003, p=0.0051, respectively). Therefore, we concluded that ZDG may treat YDH syndrome by regulating immune response, especially complement and coagulation cascades, and L‐selectin, CRP and Azgp1 may act as potential biomarkers to evaluate the therapeutic efficacy of ZDG. Our work provides important experimental basis to understand the mechanism of ZDG for treating YDH syndrome, and provide potential serum biomarkers for evaluating therapeutic efficacy of ZDG. Support or Funding Information This work was supported by grants from National Basic Research Program of China (No.2014CB543002), National Natural Science Foundation of China (No.81573709, No.81273882).

Pulmonary tuberculosis (TB) is among the diseases with the highest morbidity and mortality worldwide. Effective diagnostic methods for TB are lacking. In this study, we investigated long non-coding RNAs (lncRNAs) in plasma using microarray and the potential diagnostic value of lncRNAs for TB. We found a total of 163 up-regulated lncRNAs and 348 down-regulated lncRNAs. Gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) and coding-noncoding co-expression (CNC) analyses showed that functions of differentially expressed lncRNAs were mainly enriched in the regulation of alpha-beta T cell activation and the T cell receptor signalling pathway. Four differentially expressed lncRNAs, NR_038221 (fold change = 3.79, P < 0.01), NR_003142 (fold change = 1.69, P < 0.05), ENST00000570366 (fold change = 3.04, P < 0.05), and ENST00000422183 (fold change = 2.11, P < 0.001), were verified using RT-qPCR. Among those, NR_038221, NR_003142, and ENST00000570366 were found to be up-regulated, while ENST00000422183 was down-regulated. The value of the area under the curve (AUC) for the diagnostic model consisting of the four lncRNAs was 0.845 (sensitivity = 79.2%, specificity = 75%). We further predicted 85 mRNAs and 404 miRNAs that potentially interact with these lncRNAs. Our study revealed the potential value of lncRNAs as biomarkers for early diagnosis of TB and the underlying mechanisms of these abnormally expressed lncRNAs in the pathogenesis of TB.

基础医学创新思维教学体系(新教学体系)是实施国家培养从事医学基础研究创新人才教学目标的一套教学方法体系.新教学体系以培养、拓展被教学者创造性思维为教学目的,以传授非技术性知识和非结构性知识为教学内容,在教学实践中积极倡导被教学者的主体性与个性.新教学体系教学层次高于传统医学精英教学方法体系,但不排斥传统以培养医学精英为目的的教学方法体系.将新教学体系与传统医学精英教学体系进行合理衔接、系统化处理,有望把我国的基础医学教学水平推进到一个更高、更新的层面.

Objective:To explore the therapeutic mechanism of Zhibai Dihuang Granule ( ZDG) in Yin-deficiency-heat ( YDH) syndrome.Methods:ITRAQ-2DLC-MS/MS technique was applied to screen the differentially expressed serum proteins between ZDG treated rats and YDH syndrome rats.The serum protein profiles were analyzed by bioinformatics tools,such as Gene Ontology (GO) database,Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway database.Compared with normal rats,47 differen-tially expressed proteins were shown in the serum of YDH syndrome rats,including 20 up-regulated proteins and 27 down-regulated proteins.Nineteen proteins returned to normal after ZDG treatment,which mostly involved in immune reaction and metabolism.Complement C4 (C4),coagulation factor X (F10),and Src kinase-associated phosphoprotein 2 (Skap2) participated in immune response,while L-lactate dehydrogenase A chain (Ldha),Fructose-bisphosphate aldolase A (Aldoa),Arachidonate 12-lipoxyge-nase (Alox12),Rho GDP-dissociation inhibitor 1 (Arhgdia),and Myosin light polypeptide 6 (Myl6) mainly related to metabolic process.Conclusion:YDH syndrome is highly associated with the disturbance of immune function and metabolism,and ZDG treats YDH syndrome mostly through regulating immune activity and metabolic process.

This study aims to explore the effect of Zhibai Dihuang Granule (ZDG) on anti‐inflammatory proteins in the serum of Yin deficiency heat syndrome rats. The Yin deficiency syndrome rat model was established by triiodothyronine, and differentially expressed proteins in the serum of rats were analyzed by iTRAQ. The levels of anti‐inflammatory proteins such as Gelsolin (Gsn), Plasminogen (Plg), Fibrinogen alpha chain (Fga), and Thymosin beta‐4 (T‐β4) were significantly increased in the ZDG treated model rats, compared with the control rats. However, the level of Coagulation factor XIII A chain was significantly decreased in the ZDG treated group. Gsn has been found to remove the damaged tissue and release actin, thereby inhibiting cell apoptosis. Gsn was significantly increased in the ZDG treated group, indicating that ZDG may affect Gsn to inhibit cell apoptosis. Colla2 and Plg have been shown to regulate the blood rheology in the inflammatory response. Colla2 was significantly decreased and Plg wes significantly increased in the ZDG treated group, suggesting that ZDG can regulate the blood rheology of the inflammatory response by down‐regulating the Colla2 and up‐regulating Plg. Gsn and T‐β4 have been shown to promote the healing of inflammatory damage by regulating cytokines. Gsn and T‐β4 were significantly increased in the ZDG treated group, indicating that ZDG could inhibit apoptosis by regulating cytokines. Conclusion ZDP can treat Yin deficiency heat syndrome by removing damaged tissue, releasing actin and lipopolysaccharide, inhibiting cell apoptosis, regulating inflammatory factors and the blood rheology of inflammation, and promoting the healing of inflammatory damage. Support or Funding Information This work was supported by grants from the National Basic Research Program of China (No.2014CB543002), the National Natural Science Foundation of China (No.81573709, No.81273882)

阴虚"上火"属于中医阴虚证候,口腔炎性反应反复发作是其标志性临床体征.探究阴虚"上火"本质特征对于治疗具有十分重要意义.有研究表明,三碘甲腺原氨酸(triiodo-thyronine;T3)处理大鼠,能模拟甲亢多汗、怕热、急躁、神经过敏、震颤、心输出量增加和心率加快等类似于阴虚症状,形成大鼠阴虚模型.本实验通过腹腔注射T3处理SD大鼠建立阴虚模型,并在此基础上叠加口腔牙龈炎模型,建立阴虚"上火"大鼠模型,以iTRAQ-2DLC- MS/MS蛋白组筛选鉴定技术与生物学信息手段从血清蛋白层面探究阴虚"上火"的本质特征,为阴虚"上火"研究与治疗提供生物学依据.

Yin-deficiency-heat (YDH) syndrome is a concept in Traditional Chinese Medicine (TCM) for describing subhealth status. However, there are few efficient diagnostic methods available for confirming YDH syndrome. To explore the novel method for diagnosing YDH syndrome, we applied iTRAQ to observe the serum protein profiles in YDH syndrome rats and confirmed protein levels by ELISA. A total of 92 differentially expressed proteins (63 upregulated proteins and 29 downregulated proteins), which were mainly involved in complement and coagulation cascades and glucose metabolism pathway, were identified by the proteomic experiments. Kininogen 1 (KNG1) was significantly increased (p < 0.0001), while apolipoprotein C-III (APOC3, p < 0.005) and paraoxonase 1 (PON1, p < 0.001) were significantly decreased in the serum of YDH syndrome rats. The combination of KNG1, APOC3, and PON1 constituted a diagnostic model with 100.0% sensitivity and 85.0% specificity. The results indicated that KNG1, APOC3, and PON1 may act as potential biomarkers for diagnosing YDH syndrome. KNG1 may regulate cytokines and chemokines release in YDH syndrome, and the low levels of PON1 and APOC3 may increase oxidative stress and lipolysis in YDH syndrome, respectively. Our work provides a novel method for YDH syndrome diagnosis and also provides valuable experimental basis to understand the molecular mechanism of YDH syndrome.

The epidemic of pulmonary tuberculosis (TB), especially multidrug-resistance tuberculosis (MDR-TB) presented a major challenge for TB treatment today.We performed iTRAQ labeling coupled with two-dimensional liquid chromatography-tandem mass spectrometry (2D LC-MS/MS) and Solexa sequencing among MDR-TB patients, drug-sensitive tuberculosis (DS-TB) patients, and healthy controls.A total of 50 differentially expressed proteins and 43 differentially expressed miRNAs (fold change >1.50 or <0.60, P<0.05) were identified in the MDR-TB patients compared to both DS-TB patients and healthy controls.We found that 22.00% of differentially expressed proteins and 32.56% of differentially expressed miRNAs were related, and could construct a network mainly in complement and coagulation cascades.Significant differences in CD44 antigen (CD44), coagulation factor XI (F11), kininogen-1 (KNG1), miR-4433b-5p, miR-424-5p, and miR-199b-5p were found among MDR-TB patients, DS-TB patients and healthy controls (P<0.05)by enzyme-linked immunosorbent assay (ELISA) and SYBR green qRT-PCR validation.A strong negative correlation, consistent with the target gene prediction, was found between miR-199b-5p and KNG1 (r=-0.232,P=0.017).Moreover, we established the MDR-TB diagnostic model based on five biomarkers (CD44, KNG1, miR-4433b-5p, miR-424-5p, and miR-199b-5p).Our study proposes potential biomarkers for MDR-TB diagnosis, and also provides a new experimental basis to understand the pathogenesis of MDR-TB.

This study aimed to observe the expression profiles of serum proteins and to discover potential biomarkers in Yin‐deficiency‐heat (YDH) syndrome rats induced by hot and pungent Chinese herbs. Totally 50 Sprague–Dawley rats were randomly divided into the normal group and the YDH model group. The concentrated decoction of three Chinese herbs such as aconite root, dried ginger, and cinnamon were applied to establish YDH model. The signs and symptoms of rats were observed dynamically. All rats were sacrificed at the end of the 4th week and serum was collected by using carotid artery retaining needle. Serum protein profiles in YDH rats and normal rats were investigated by iTRAQ‐coupled 2DLC‐MS/MS technique. Compared with the normal group, rats in the YDH model group showed obvious YDH symptoms, including dry hair, restlessness, and weight loss. Rats in the YDH model group gained weight more slowly (p<0.01) than did those of the normal group, and the thymic index decreased significantly (p<0.001) in the YDH model group. When setting quantification ratio of >1.25 as up‐regulated threshold and <0.8 as down‐regulated threshold, a total of 165 differentially expressed proteins (110 up‐regulated proteins and 55 down‐regulated proteins) were identified in the serum of YDH syndrome rats. Significant differences in protein Kininogen (KNG), Apolipoprotein C‐III (ApoC‐III) and Paraoxonase1 (PON1) were found between the YDH model group and the normal group by ELISA. Our work provided valuable insights into the changes of the protein expression in YDH syndrome rats, and identified 3 differentially expressed proteins (KNG, ApoC‐III and PON1) that can be used as potential biomarkers for diagnosing YDH syndrome.Support or Funding InformationThis work was supported by grants from National Basic Research Program of China (No.2014CB543002), National Special Sci‐Tech Projects (No. 2012ZX10005001‐006), and National Natural Science Foundation of China (No.81273882, NO. 81573709).

OBJECTIVE To observe the preliminary action of phosphodiesterase 4B(PDE4B) in bleomycin-induced pulmonary fibrosis in mice.METHODS Pulmonary fibrosis in mice was induced by intratracheal instillation of belomycin 2.5 mg·kg-1.On the 3rd,7th,14th,21st and 28th day after instillation,bronchoalveolar lavage and lung tissue specimens were collected.The total number of white blood cells count and differential count in bronchoalveolar lavage fluid(BALF) were recorded.Transforming growth factor-β1(TGF-β1),macrophage inflammatory protein-2(MIP-2),interleukin-6(IL-6) and IL-1β levels in BALF were determined by ELISA.Collagen content and myeloperoxidase(MPO) activity were determined by assay kits.PDE4B mRNA expression in mouse lung tissue was detected by quantitative real-time PCR.Immunohistochemistry was used to detect the PDE4B protein expression and distribution in lung tissue.RESULTS After airway instillation,the bleomycin-induced pulmonary fibrosis continued to develop,reaching the peak on the 28th day and showing clearly lung fibrosis.After modeling,the total number of leukocytes in BALF,IL-1β and IL-6 reached the peak on the 3rd day,and the peak value was 22.0,2.0 and 2.8 fold,respectively,compared with normal control.MPO and TGF-β1 reached the peak on the 7th day,which was 1.9 and 5.5 fold,respectively,compared with normal control.While the collagen content,MIP-2 and PDE4B mRNA expression from the modeling continued to increase,the peak value was 1.6,2.7 and 2.6 fold,respectively,compared with normal control.Immunohistochemical results showed that the PDE4B protein was distributed in the inflammatory cells and fibrosis of the lung tissue.CONCLUSION PDE4B plays an important role in pulmonary fibrosis,and may be a specific drug target.

OBJECTIVE To study the effect of AICAR on uterine smooth muscle of pregnant and nonpregnant rats.METHODS The action of AICAR and dibutyryl-cAMP on the contractile function of isolated myometrium of pregnant and nonpregnant Sprague-Dawley rats was investigated.AICAR or dibutyryl-cAMP was used at cumulative doses from 10-9 to 10-4 mol·L-1 for myometrium of pregnant rats,and at 10-5 mol·L-1 for myometrium of nonpregnant rats.The amplitude of the contractility of myometruim was recorded.RESULTS For the myometrium of pregnant or nonpregnant rats,AICAR and dibutyryl-cAMP exerted strong inhibitory action on the contractile function in dose-dependent manners.For the myometrium of pregnant rats,AICAR or dibutyryl-cAMP showed inhibitory effect at a low concentration of 10-7 mol·L-1.AICAR and dibutyryl-cAMP also caused significant inhibition in myometrium from nonpregnant rats.CONCLUSION AICAR exerted strong inhibitory action on the contractile function of myometrium from pregnant or nonpregnant rats,suggesting that AICAR may have inhibitory effect on spontaneous activity of myometrium of human.

Knuckle pads and camptodactyly are overlapping symptoms associated with many genetic and environmental factors. To the best of our knowledge, all reported cases of epidermolytic palmoplantar keratoderma (EPPK) with knuckle pads have been without accompanying camptodactyly. We here report a novel KRT9 mutation-EPPK family with combined knuckle pads and camptodactyly. All the EPPK-affected individuals in this southern Chinese pedigree suffered severe diffuse palmar and plantar hyperkeratosis including hyperhidrosis and cuticle splitting: 3 females presented EPPK only, 8 adult males had notably severe knuckle pads and camptodactyly as well as EPPK, and one 6-year-old boy manifested EPPK with knuckle pads. Haplotype analysis excluded the known candidate loci for camptodactyly and/or knuckle pad-like phenotypes on chromosomes 13q12, 3q11.2-q13.12, 1q24-q25, 4p16.3 and 16q11.1-q22, while only the markers D17S1787 and D17S579 flanking KRT9 showed co-segregation with EPPK. Then a novel c.T1373C (p.L458P) mutation within the sixth exon of KRT9 was validated, and this mutation presented a more severe pathogenicity than the previously reported p.L458F. We speculated that KRT9 plays a complicated role in the genesis of EPPK with knuckle pads and camptodactyly, which needs to be further investigated.

In the present study, we investigated the effect of classic PDE4 inhibitor rolipram and novel PDE4 inhibitor ZL-n-91 on LPS-induced acute lung injury (ALI) in mice and its mechanism. ALI was induced in ICR mice by instilling intratracheally with LPS, and mice were divided into seven groups: control (Saline), LPS group, ZL-n-91 (3μg, 10μg, and 30μgkg−1, ip), Rolipram (1.0mgkg−1, ip) and dexamethasone (0.5mgkg−1, ip). After the 6h of instilling intratracheally with LPS in mice, total leukocyte number, neutrophil number and protein content in BALF increased rapidly, a large number of neutrophil infiltration around the pulmonary vessel and airway, the lung wet weight/dry weight (w/d)ratio raised significantly. MPO activity, TNF-α level and cAMP-PDE, PDE4 activity in lung homogenate raised significantly. PaO2, PaCO2 and PH value in peripheral arterial blood also changed obviously, PaO2 and PH value dropped slightly and PaCO2 increased significantly in LPS group. ZL-n-91 (3μg, 10μg, 30μgkg−1) dose-dependently reduced the total leukocyte number, neutrophil number and total protein content in BALF, MPO activity, TNF-α level and cAMP-PDE, PDE4 activity in lung homogenate, but the effect of ZL-n-91 in pathological changes and lung wet w/d ratio is slight; Rol and Dex significantly reduced lung wet w/d ratio and improved pathological changes, neutrophil around the pulmonary vessel and airway significantly reduced, symptoms of lung edema relieved; The PH value, PaO2 and PaCO2 in ZL-n-91 high dosage group and Rol group had changes, but there was no significant difference compared with LPS group or saline group; After the administration, the righting reflex recovery time significantly shorten in every group of ZL-n-91. the righting reflex recovery time of Rol group was similar with ZL-n-91 30μgkg−1 group, while Dex group was similar with saline group. The present study confirms that the inhibitory effect of ZL-n-91(30μgkg−1) on the inflammatory reactivity, including inhibition of inflammatory cell and protein exudation, MPO and PDE4 activity, improvement of the blood gas, those effects were equivalent with rolipram 1mgkg−1, and suggested that ZL-n-91 was stronger than rolipram in PDE4 inhibition. So we speculated that ZL-n-91 may have stronger therapeutic potential for treatment of inflammatory disease than rolipram, meantime have stronger nervous system effect than rolipram.