Zarzio Ameliorates Non-alcoholic Fatty Liver Induced By a High Fat Diet in a Rat Experimental Model: A Histological and Immunohistochemical Study

SUMMARY: The therapeutic effect of a granulocyte-colony stimulating factor (G-CSF) biosimilar drug, zarzio, on non-alcoholic fatty liver disease (NAFLD) in a rat model was investigated in this study. Thirty-two rats were randomly divided into four groups. Groups I and II were fed a standard laboratory diet, whereas groups III and IV were fed a high fat diet (HFD) for 14 weeks. After 12 weeks of feeding, groups I and III were administered normal saline, and groups II and IV were intraperitoneally administered zarzio (200 mg/kg/day) for two consecutive weeks. Hematoxylin-eosin (H&E) staining was used to assess hepatic and pancreatic morphology in all groups, oil red O (ORO) staining for lipid accumulation, Masson's staining for fibrosis, and immunohistochemistry assay for hepatic protein expression of insulin receptor substrate 1 (IRS1), nuclear factor erythroid 2-related factor 2 (Nrf2), tumour necrosis factor alpha (TNF-α) and pancreatic caspase-3. The NAFLD rats (group III) developed hepatic steatosis with increased lipid accumulation, perisinusoidal fibrosis, upregulated IRS1, TNF-α (all P<0.05) without a significant increase in Nrf2 protein expression compared with normal control. In comparison, model rats treated with zarzio (group IV) showed significant rejuvenation of the hepatic architecture, reduction of fat accumulation, and fibrosis. This was accompanied by the upregulation of Nrf2, downregulation of IRS1 and TNF-α protein expression (all P<0.05). No correlation was detected between NAFLD and non-alcoholic fatty pancreas disease (NAFPD). However, the pancreatic β-cells in group III showed increased caspase-3 expression, which was decreased (P<0.05) in group IV. In conclusion, zarzio ameliorates NAFLD by improving the antioxidant capacity of liver cells, reducing hepatic IRS1, TNF-α protein expression and pancreatic β-cells apoptosis, suggesting that zarzio could be used as a potential therapy for NAFLD.

En este estudio se investigó el efecto terapéutico de un fármaco biosimilar del factor estimulante de colonias de granulocitos (G-CSF), zarzio, sobre la enfermedaddel hígado graso no alcohólico (NAFLD) en un modelo de rata. Treinta y dos ratas se dividieron aleatoriamente en cuatro grupos. Los grupos I y II fueron alimentados con una dieta estándar de laboratorio, mientras que los grupos III y IV fueron alimentados con una dieta alta en grasas (HFD) durante 14 semanas. Después de 12 semanas de alimentación, a los grupos I y III se les administró solución salina normal, y a los grupos II y IV se les administró zarzio por vía intraperitoneal (200 mg/kg/ día) durante dos semanas consecutivas. Se utilizó tinción de hematoxilina-eosina (H&E) para evaluar la morfología hepática y pancreática en todos los grupos, tinción con rojo aceite O (ORO) para la acumulación de lípidos, tinción de Masson para la fibrosis y ensayo de inmunohistoquímica para la expresión de la proteína hepática del sustrato 1 del receptor de insulina (IRS1), factor nuclear eritroide 2 relacionado con el factor 2 (Nrf2), factor de necrosis tumoral alfa (TNF-α) y caspasa-3 pancreática. Las ratas NAFLD (grupo III) desarrollaron esteatosis hepática con aumento de la acumulación de lípidos, fibrosis perisinusoidal, IRS1 y TNF-α regulados positivamente (todos P <0,05) sin un aumento significativo en la expresión de la proteína Nrf2 en comparación con el control normal. En comparación, las ratas modelo tratadas con zarzio (grupo IV) mostraron un rejuvenecimiento significativo de la arquitectura hepática, una reducción de la acumulación de grasa y fibrosis. Esto estuvo acompañado por la regulación positiva de Nrf2, la regulación negativa de la expresión de la proteína IRS1 y TNF-α (todas P <0,05). No se detectó correlación entre NAFLD y la enfermedad del páncreas graso no alcohólico (NAFPD). Sin embargo, las células β pancreáticas en el grupo III mostraron una mayor expresión de caspasa-3, que disminuyó (P <0,05) en el grupo IV. En conclusión, zarzio mejora la NAFLD al mejorar la capacidad antioxidante de las células hepáticas, reduciendo el IRS1 hepático, la expresión de la proteína TNF-α y la apoptosis de las células β pancreáticas, lo que sugiere que zarzio podría usarse como una terapia potencial para la NAFLD.

Effects of volatile oil from Acori graminei Rhizoma on glial fibrillary acidic protein, c-Jun N-terminal protein kinase and tumor necrosis factor-α expressions in the spinal cord dorsal horn of the inflammatory pain rats / 解剖学报

Objective To investigate the influence of volatile oil from Acori graminei Rhizoma (VOA) on expressions of glial fibrillary acidic protein (GFAP), c-Jun N-terminal protein kainse (JNK) and tumour necrosis factor-α (TNF-α) in the spinal cord dorsal horn of imflammatory pain rats. Methods Totally 36 male SD rats were randomly divided into control group (control), sham-operated group (sham), complete Freund' s adjuvant group (CFA), 5 g/(kg·d) low dose VOA+CFA group (VOA-L+CFA), 10 g/(kg·d) medium dose VOA + CFA group (VOA-M+CFA) and 20 g/(kg·d) high dose VOA + CFA group (VOA-H+CFA). All animals were sacrificed immediately after continuous gavage administration for 22 days. The expressions of GFAP, JNK and TNF-α in the spinal cord dorsal horn of rats in each group were detected by immunofluorescence and Western blotting methods. Results The present results showed that the positive expressions of GFAP, JNK and TNF-α in the spinal cord dorsal horn of rats increased significantly in the CFA group, when compared to the control and sham groups (P < 0. 01). The expressions of GFAP, JNK and TNF-α in the spinal cord dorsal horn of rats with VOA treatment reduced in the dose-dependent manner, when compared to the CFA group, the positive expressions of GFAP, JNK and TNF-α reduced significantly in the dorsal horn of the spinal cord of the VOA-H+CFA group (P<0. 05, P<0. 01). Conclusion VOA reduces the expressions of GFAP, JNK and TNF-α in the spinal cord dorsal horn of rats of CFA-induced inflammatory pain.

Yes-associated protein ameliorates tumor necrosis factor-α-induced epithelial barrier dysfunction by affecting the F-actin / 中国小儿急救医学

Objective:To investigate the protective role of Yes-associated protein(YAP)in intestinal epithelial barrier injury.Methods:The intestinal epithelial barrier model was established by culturing human colorectal adenocarcinoma cell line Caco-2 cells, which were divided into four groups: control group, Caco-2 monolayers did not receive any treatment; recombinant human tumor necrosis factor-α(rhTNF-α)group, 100 μg/L of rhTNF-α was added to Caco-2 monolayers; vector+ rhTNF-α group, Caco-2 monolayers were first added with control plasmid pcDNA3.1-vector, and 100 μg/L rhTNF-α was added 24 hours later; YAP+ rhTNF-α group, Caco-2 cells with barrier construction were first added with pcDNA3.1-YAP, and 100 μg/L rhTNF-α was added 24 hours later.Realtime-PCR and Western blot were used to evaluate YAP mRNA and protein expression level.Epithelial permeability was assayed by trans-epithelial electrical resistance(TEER)and fluorescein isothiocyanate-dextran 40(FD-40 flu). Cellular distribution of F-actin was assayed by immunofluorescence staining.Results:Compared with control group[(607.3±29.3)Ω·cm 2], TEER of rhTNF-α group[(265.3±32.7)Ω·cm 2] decreased, while TEER of YAP+ rhTNF-α group[(387.0±18.7)Ω·cm 2]increased compared with rhTNF-α group, the differences were statistically significant( P<0.001). The FD-40 flux of rhTNF-α(22.7%±0.5%) group was higher than that of the control group(6.3%±0.9%), while the FD-40 flux of Yap + rhTNF-α group(12.2%±0.8%) was lower than that of rhTNF-α group, the differences were statistically significant( P<0.001). Immunofluorescence staining showed that compared with the control group, the cytoskeletal F-actin fiber dense spot decreased in rhTNF-α group, and some cells showed obvious trans-cellular stress fiber structure, while the peripheral actin band was clear in YAP+ rhTNF-α group, and the intracellular stress fiber decreased.YAP+ TNF-α group appeared as a clear, peripheral actin ribbon with a decrease in cytoplasmic stress fibres. Conclusion:YAP overexpression significantly inhibits TNF-α induced decline of TEER, and increases of FD-40 flux and F-actin rearrangement of Caco-2.YAP could ameliorate TNF-α induced intestinal epithelial barrier injury by regulating cytoskeleton F-actin.

A study on keloid fibroblast induced THP-1 derived macrophage polarization and inflammatory factor TNF-αexpression and its feedback effect / 中华医学美学美容杂志

Objective:To investigate the effect of keloid fibroblasts on the polarization and expression of inflammatory factors of M0 macrophages and possible mechanisms, and provide theoretical basis for new targets for keloid therapy.Methods:Keloids, normal skin tissues and paraffin specimens from patients undergoing plastic surgery in the First Affiliated Hospital of Sun Yat-sen University from November 2020 to September 2021 were collected, and fibroblasts of keloids and normal skins were isolated and co-cultured with M0 cells formed form THP-1 by phorbol ester (PMA)-stimulation to detect the expression of macrophage polarization markers and cytokines. Besides, keloid fibroblasts were treated with exogenous tumor necrosis factor-α(TNF-α) to detect its effect on the proliferation and extracellular matrix expression.Results:Macrophages were dominated by CD163 + (M2) in keloid tissues. Moreover, M0 cells expressed more TNF-α when co-cultured with keloid fibroblasts, compared with those with normal skin fibroblasts, in which, the positive staining rates of TNF-α were 19.32% and 29.52% respectively by flow cytometry. Furthermore, the proliferation was promoted and the expression of extracellular matrix proteins (COL3A1 and FN1)and Vimentin were upregulated in keloid fibroblasts under TNF-α stimulation. However, there was no significant difference in the expression of polarization surface markers CD86 and CD163 in macrophages, when co-cultured with keloid fibroblasts or normal skin fibroblasts. Conclusions:Keloid fibroblasts promote the expression of TNF-α in macrophages, which in turn promotes the proliferation and extracellular matrix secretion of keloid fibroblasts.

Huangqin Decoction Attenuates DSS-Induced Mucosal Damage and Promotes Epithelial Repair via Inhibiting TNF-α-Induced NF-κB Activation / 中国结合医学杂志

OBJECTIVE@#To investigate the protective effect of Chinese herbal formula Huangqin Decoction (HQD) on ulcerative colitis mouse model induced by dextran sulphate sodium (DSS) and human intestinal epithelial cell injury induced by tumour necrosis factor-α (TNF-α).@*METHODS@#In vivo, 30 male C57BL/6 mice were divided into 5 groups using a random number table (n=6 per group), including control, DSS, 5-aminosalicylic acid (5-ASA), HQD low- (HQD-L) and high-dose (HQD-H) groups. The colitis mouse model was established by 3% (w/v) DSS water for 5 days. Meanwhile, mice in the HQD-L, HQD-H and 5-ASA groups were administrated with 100, 200 mg/kg HQD or 100 mg/kg 5-ASA, respectively, once daily by gavage. After 9 days of administration, the body weight, disease activity index (DAI) score and colon length of mice were measured, the pathological changes of colons were analyzed by hematoxylin-eosin staining (HE) staining, and the levels of serum interleukin (IL)-6, IL-1β and TNF-α were measured by enzyme linked immunosorbent assay. In vitro, the human colon epithelial normal cells (FHC cells) were exposed to HQD (0.6 mg/mL) for 12 h and then treated with TNF-α (10 ng/mL) for 24 h. The tight junction (TJ) protein expression levels of Claudin-4 and Occludin, and the protein phosphorylation levels of p65 and inhibitor of nuclear factor kappaB (NF-κB)-α (IκBα) were measured by Western blot.@*RESULTS@#In vivo, compared with the DSS group, HQD-H treatment attenuated the weight loss and reduced DAI score of mice on the 8th day (P<0.05). Moreover, HQD-H treatment ameliorated the colon shortening in the DSS-induced colitis mice (P<0.05). HE staining showed HQD attenuated the pathological changes of colitis mice, and the histological scores of HQD-H and 5-ASA groups were significantly decreased compared with the DSS group (P<0.05). Meanwhile, HQD-H and 5-ASA significantly decreased the serum IL-1β, IL-6 and TNF-α levels of mice (P<0.05). In vitro experiments showed that HQD up-regulated Occludin and Claudin-4 protein expressions and inhibited p-p65 and p-IκBα levels in FHC cells compared with the TNF-α group (P<0.05).@*CONCLUSION@#HQD significantly relieved the symptoms in DSS-induced colitis mice by inhibiting pro-inflammatory cytokines expression and maintained the homeostasis of TJ protein in FHC cells by suppressing TNF-α-induced NF-κB activation.

Relationship between serum complement C1q/tumor necrosis factor related protein 3 level and carotid atherosclerosis in patients with diabetes mellitus complicated with fatty liver / 中国综合临床

Objective:To investigate the correlation between serum complement C1q/tumor necrosis factor associated protein 3 (CTRP3) and carotid atherosclerosis in patients with type 2 diabetes mellitus (T2DM) and nonalcoholic fatty liver disease (NAFLD).Methods:From January 2018 to December 2019, 111 T2DM patients hospitalized in the Endocrinology Department of Nantong Third People ′s Hospital Affiliated to Nantong University, and 30 healthy physical examiners in the physical examination center of Nantong Third People 's Hospital Affiliated to Nantong University in the same period were selected. Thirty cases of healthy physical examination were the control group, 111 cases of T2DM were divided into 52 cases of T2DM group and 59 cases of T2DM+NAFLD group according to whether they were combined with NAFLD. The cross-sectional study method was used to collect the relevant clinical data of three groups. The comparison data between multiple groups conformed to the normal distribution and the variance was uniform. One way ANOVA was used. SNK- q test was used for pairwise comparison, χ2 test for qualitative data comparison. The correlation between carotid intima-media thickness (IMT) and influencing factors was analyzed by partial correlation analysis, and the influencing factors of carotid IMT were analyzed by multi factor linear regression. Results:In the control group, T2DM group and T2DM+NAFLD group, body mass index (BMI) (23.65±2.81), (25.52±3.12), (24.90±2.94) kg/m 2,systolic blood pressure (119.43±15.81), (130.63±10.20), (139.37±14.11) mmHg, diastolic blood pressure (72.93±9.74), (73.40±9.44), (77.97±10.00) mmHg, and fasting blood glucose (5.12±0.77), (9.78±1.37), (9.24±1.46) mmol/L,glycosylated hemoglobin (HbA1c) (4.87±1.43)%, (7.99±1.10)%, (8.56±1.29)%,homeostasis model assessment of insulin resistance (HOMA-IR)(1.56±0.37),(2.80±1.00), (3.47±0.94), high density lipoprotein cholesterol (HDL-C) (1.52±0.34),(1.23±0.31), (1.22±0.31) mmol/L,low density lipoprotein cholesterol (LDL-C) (2.41±0.53), (2.73±0.61), (2.93±0.59) mmol/L, CTRP3 (292.93±68.54), (241.69±61.01), (150.80±56.67) μg/L, the difference between groups were statistically significant ( F=3.712,23.023,4.074,134.285,90.818,47.105,10.139,7.941,60.035,all P<0.05). Pairwise comparison shows that the systolic blood pressure, diastolic blood pressure, HbA1c and HOMA-IR in T2DM+NAFLD group were higher than those in control group and T2DM group,and CTRP3 was lower than those in control group and T2DM group, the difference was statistically significant (all P<0.05). BMI, fasting blood glucose, HbA1c, HOMA-IR, HDL-C and LDL-C in T2DM group were higher than those in the control group, CTRP3 was lower than that in the control group (all P<0.05). In the control group, T2DM group and T2DM+NAFLD group, IMT were (0.75±0.13), (1.11±0.17) and (1.25±0.15) cm; Crouse scores were (1.28±0.97), (3.22±1.42) and (4.54±1.22); the plaque detection rates 16.7%(5/30), 65.4%(34/52) and 78.0%(46/59), and there were significant differences between the two groups ( F=105.941,67.063, χ2=32.108, all P<0.001). There were significant differences between the two groups (all P<0.05). T2DM+NAFLD group was the highest, followed by T2DM group, and the control group was the lowest. Partial correlation analysis showed that carotid IMT was positively correlated with systolic blood pressure, fasting blood glucose, HbA1c, HOMA-IR, triglyceride and LDL-C ( r=0.356, 0.572, 0.575, 0.620, 0.172, 0.291, all P<0.05), and negatively correlated with HDL-C and CTRP3 ( r=-0.335, -0.675, all P<0.001). Multivariate linear regression analysis showed that HbA1c, HDL-C and ctrp3 were the influencing factors of carotid atherosclerosis ( t=2.621, -3.764, -7.280, all P<0.05) Conclusion:Serum CTRP3 is associated with carotid atherosclerosis in T2DM patients with NAFLD,and may have a protective effect on vascular lesions in T2DM patients with NAFLD.

Herbal Medicine from Single Clove Garlic Oil Extract Ameliorates Hepatic Steatosis and Oxidative Status in High Fat Diet Mice

@#Introduction: High fat diet (HFD) can cause lipid accumulation and contribute to various metabolic disorders. Single clove garlic oil (SCGO) has advantages over regular garlic due to its higher amounts of organosulfide compounds in particular. This study aimed to determine the ability of SCGO extract to ameliorate hepatic steatosis and improve oxidative status by modulating expression of tumour necrosis factor α and superoxide dismutase in mice fed a HFD. Methods: Twenty-four adult male Balb/C mice were divided into six groups: i) normal diet; ii) positive control diet; iii) negative control diet; and iv) HFD with SCGO at 12.5 mg/kg body weight (mg/kg BW); v) HFD with SCGO at 25 mg/kg BW, vi) HFD with SCGO at 50 mg/kg BW. Liver weight and morphology, spleen weight, serum levels of superoxide dismutase (SOD) and tumour necrosis factor α (TNF-α), TNF-α expression in the aorta and lipid profiles were assessed at the end of the experimental period. Results: SCGO treatment was associated with significant decreases in liver and spleen weight as well as amelioration of hepatic steatosis. SCGO treatment also decreased TNF-α levels and expression. Serum levels of SOD in the SCGO groups were significantly increased compared with the negative control group. Lipid profiles were improved in the SCGO treatment groups compared with the negative control group. Conclusion: SCGO as an herbal medicine could be an effective treatment for degenerative disorders caused by HFD.

Correlation of Small Dense Low Density Lipoprotein, Tumour Necrosis Factor - alpha with Liver Enzymes in Chronic Hepatitis B Patients

Aim:This study investigated the relationship between small dense low density lipoprotein (sdLDL), tumour necrosis factor-alpha (TNF-α), aspartate aminotransferase (AST), alanine aminotransferase(ALT) and alkaline phosphatase (ALP) in chronic hepatitis B patients.Duration of Study: June2018-March 2019.Subjects and Methods:Sixty (60) participants were recruited for this cross sectional study. They comprised thirty (30) clinically diagnosed chronic hepatitis B virus (HBV) infected patients attending clinic at a tertiary hospital in Osogbo, Osun state, Nigeria. Thirty (30) apparently healthy volunteers were recruited as control subjects after fulfilling the inclusion criteria. Anthropometric measurements were performed using standard method. About 6mL of venous blood was collected from each study participant,serum was extracted and kept at -80oC until time of analysis. Small dense LDL, TNF-α, AST, ALT and ALP were determined using enzymelinked immunosorbent assay and colorimetric method as appropriate. Data analysis was doneusing Student’s t-test for Original ResearchArticle comparison of variables and Pearson’s correlation was used to determine the relationship between variables. P–value less than 0.05 was considered significant. Results:SdLDL, TNF-α, AST and ALT were significantly elevated in HBV patients when compared with the control subjects (P<0.05). SdLDL had a significant positive correlation with TNF-α (P=0.03), AST (P=0.01), ALT (P=0.00). TNF-α had a significant positive correlation with AST (P=0.02) and ALT (P=0.00).Conclusion:This study revealed a noteworthy positive relationship between sdLDL, TNF-α and hepatic aminotransferases in chronic hepatitis B patients

Genetic and expression changes in TNF-α as a risk factor for rheumatoid arthritis pathogenesis in northeast India

Antitumour necrosis factor-alpha (TNF-α) therapy is used as a clinical intervention for rheumatoid arthritis (RA) but differences exist in response to the treatment which makes the candidature of the screening of TNF-α alteration(s) at genetic and expression levels an important agenda prior to treatment. This study aims to determine the associative role of TNF-α –308G/A polymorphism and differential expression of TNF-α in the pathogenesis of RA. A case–control study where a total of 126 RA patients were enrolled based on ACR-EULAR (2010) criteria, along with 160 community matched age and sex controls over a period of three years. The differential expression level of TNF-α mRNA and protein level was studied and TNF-α –308G/A polymorphism was screened by T-ARMS PCR assay. All statistical analysis was performed using SPSS software. mRNA expression level of TNF-α was upregulated in RA cases (avg. 15.85 ± 9.52 fold) compared to control. TNF-α protein level was found to be higher in RA cases (28.62±7.17 pg/mL) compared to control (23.14±6.91 pg/mL). TNF-α –308 variant GA genotype was higher in RA (46.03%) than in control (25%). The presence of TNF-α –308 variant A allele was associated with increased risk of RA susceptibility (odds ratio (OR) = 2.559 at 95% confidence interval (CI), P< 0.001) but not severity (OR = 1.617 at 95% CI, P = 0.571). The presence of –308 variant genotype was associated with a higher TNF-α mRNA and protein expression. The presence of TNF-α –308A allele is associated with increased risk of RA susceptibility and differential TNF-α expression, and has prognostic significance. Association of higher TNF-α pro-inflammatory cytokine levels with northeast Indian patients makes them suitable subjects for anti-TNF-α therapy.

Clinical characteristics of autoinflammatory diseases: a retrospective analysis in a single centre over a 5-year period / 中国实用内科杂志

OBJECTIVE: This study is aimed to evaluate the features in patients with autoinflammatory diseases and to assess the applicability of the international clinical diagnostic criteria for autoinflammatory diseases in these patients. METHODS: We retrospectively reviewed clinical data patients with autoinflammatory diseases in Peking University First Hospital within 5 year.RESULTS: Totally 50 patients were included. Eighteen patients experienced their first attack before 18 years of age, and 32 patients were with adult onset. The median age at onset was 25.5 years(range 1-74); 35(70%) cases experienced recurrent episodes of fever;15(30%) cases had continuous fever. Inflammatory markers were elevated in most patients during fever attack, and reduced in period with no symptoms. All of patients had one or more sequence variants(SVs). MEFV gene mutations were the most common and all SVs were heterozygous.The most frequent genotype was E148 Q(23 patients 50%).Only 5 MEFV SVs cases(10.8%)were up to the familial Mediterranean fever(FMF) Tel Hashomer clinical criteria. Totlly 7 patients were diagnosed with single-gene hereditary autoinflammatory disease. CONCLUSION: Most patients in the study didn't show typical clinical features or typical gene mutations. The international diagnostic criteria of autoinflammatory diseases is not applicable to the patients in this study.

Effects of blood glucose fluctuation on the expression of TLR4 and TNF-α in the liver of diabetic rats / 预防医学

Objective@# To investigate the effect of blood glucose fluctuation on the expression of toll-like receptor 4（TLR4）and tumour necrosis factor-α（TNF-α）in the liver of diabetic rats .@*Methods @#The adult male Sprague-Dawley（SD）rats were used to establish diabetic rats model and then they were randomly assigned to sustained hyperglycemia group（MS group,n=20） and fluctuating hyperglycemia group（MF group,n=20）. The two groups continued high-fat and high-sugar diet,while MF group alternately received intraperitoneal injection of glucose and subcutaneous injection of short-acting insulin. Another 10 SD rats were assigned to the control group. After 6 weeks,the physical signs,blood glucose,triglyceride（TG）,total cholesterol（TC）,low density lipoprotein cholesterol（LDL-C）,high density lipoprotein cholesterol（HDL-C）,alanine aminotransferase（ALT）and aspartate aminotransferase（AST）of the rats were measured;the mRNA expression of TLR4 and TNF-α in liver tissues were detected by reverse transcription-polymerase chain reaction （RT-PCR）;pathological changes of liver tissues were observed after HE staining .@*Results @#Compared with the control group,the rats in the MS group were always at the hyperglycemia status,the blood glucose of the rats in the MF group drifted between the peak and the trough. The weight growth of the rats in the MS group and MF group were slower. The levels of TC,TG and LDL-C significantly increased and the level of HDL-C significantly decreased in the MS group and MF group （all P<0.05）. The activities of ALT and AST increased both in the MS group and MF group,with the MF group increased more significantly（P<0.05）. The mRNA expression levels of TLR4 and TNF-α in liver tissues of the rats in MS group and MF group increased,with the MF group increased more significantly（P<0.05）. HE staining results showed that the liver cells of the rats in the MF group had more lipid droplet deposition,with the disordered hepatocyte line arrangement and more severe lipid droplet vacuolation. The lesion rate of the MS group and MF group were 83.30% and 100.00% .@*Conclusion@# The rats in this model showed signs of hyperglycemia complicated by dyslipidemia and liver injury. The expression of TLR4 and TNF-α increased in rats with blood glucose fluctuation,which might play a role in the aggravation of diabetic liver injury.

Effect of Shaoyaotang on Expressions of AP-1 and TNF-α in Colon of Rats with Hygrothermal Ulcerative Colitis / 中国实验方剂学杂志

Objective: To observe the effect of Shaoyaotang on mRNA and protein expressions of colon tissue activated protein-1 (AP-1) and tumor necrosis factor-α (TNF-α) of hot and humid-type intrinsic ulcerative colitis (UC) model in rats, in order to explore the mechanism of action of herbaceous peony decoction in the treatment of UC. Method: Totally 60 Wistar rats were randomly divided into blank group, model group, SASP group, and low, medium and high-dose Shaoyaotang groups. The damp-heat intrinsic UC rat model was replicated based on integrated disease and syndrome, namely, high-fat and high-sugar spicy food and immune complex method combined with 2,4,6-trinitrobenzene sulfolnic acid (TNBS) and ethanol complex method. After the successful modeling, low, medium and high-dose Shaoyaotang (6, 12, 24 g·kg-1) was given by gavage, and 1 g·kg-1 dose of salazol sulfadiazine was given to by gavage. The blank group was given constant volume normal saline for 21 d. Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) was used to detect mRNA expressions of AP-1 and TNF-α in colon tissues, and Western blot was used to detect protein expressions of AP-1 and TNF-α in colon tissues. Result: Compared with the blank group, relative mRNA and protein expressions of AP-1, TNF-α in the model group were significantly increased (Pα in the treatment groups were significantly decreased (PConclusion: Shaoyaotang can inhibit the expression of TNF-α and stimulate AP-1 protein expression in rats with damp-heat UC.

Clinical Observation on Modified Huqianwan in Treatment of Rheumatoid Arthritis Liver-kidney Yin Deficiency Syndrome / 中国实验方剂学杂志

Objective: To explore the clinical efficacy and safety of modified Huqianwan in treatment of rheumatoid arthritis (RA) liver-kidney Yin deficiency syndrome, and investigate its possible mechanism. Method: A total of 184 patients with RA liver-kidney Yin deficiency syndrome were randomly divided into Chinese medicine group (62 cases), western medicine group (57 cases) and integrated Chinese and western medicine group (65 cases) according to the digital table method. The patients in Chinese medicine group were treated with Huqianwan; the patients in western medicine group were treated with methotrexate tablets and leflunomide tablets; and the patients in integrated Chinese and western medicine group received Huqianwan+methotrexate tablets and leflunomide tablets,with a treatment course of 12 weeks in all groups. The pain visual analog scale (VAS), swelling and tenderness scores of 28 joints (DAS28), average hands grip strength, morning stiffness time and liver-kidney Yin deficiency syndrome differentiation of traditional Chinese medicine (TCM) syndrome score were compared between groups before and after treatment. The changes of erythrocyte sedimentation rate (ESR), C reactive protein (CRP), immunoglobulin (Ig) G, tumor necrosis factor-alpha (TNF-α) and rheumatoid factor (RF) were detected in all groups after treatment. Clinical efficacy, and incidence of adverse reactions such as gastrointestinal response, liver injury, leukopenia, serum glutamate oxaloacetic aminotransferase (GOT) and platelet (PLT) level changes were compared between the groups, so as to investigate the efficiency and safety of the different medicines. Result: After 12 weeks of treatment, the total clinical effective rate was 79.0%, 80.7%, and 92.3% respectively in Chinese medicine group, western medicine group, and integrated Chinese and western medicine group; the integrated Chinese and western medicine group was significantly better than the Chinese medicine group and western medicine group (PPPPConclusion: The efficacy in treating RA liver and kidney Yin deficiency syndrome shows no significant difference between modified Huqianwan and methotrexate tablets+leflunomide tablets. In the treatment of RA liver and kidney Yin deficiency syndrome, Huqianwan has fewer adverse reactions. Huqianwan combined with methotrexate tablets+leflunomide tablets is superior to that in methotrexate tablets+leflunomide tablets in treatment of RA liver-kidney Yin deficiency syndrome.

Impact of TRAIL gene knockout on regulatory T cells in mice with dextran sodium sulphate-induced experimental colitis / 中华微生物学和免疫学杂志

Objective To investigate the impact of knocking out tumor necrosis factor-related ap-optosis-inducing ligand ( TRAIL) gene ( TRAIL-/-) on colonic inflammation and regulatory T cells ( Treg) in mice with dextran sulfate sodium (DSS)-induced experimental colitis. Methods C57BL/6 mice were ran-domly assigned into four groups with 10 in each group:wild-type ( WT) control, WT colitis, TRAIL-/- con-trol and TRAIL-/- colitis. The mouse model of colitis was induced by oral administration of 3. 5％ DSS and the severity of colonic inflammation was assessed. Peripheral blood mononuclear cells ( PBMCs) and mesen-teric lymph nodes ( MLNs) were collected. The ratios of Treg cells to CD4+T cells in PBMCs were detected by flow cytometry. Expression of Treg cell-associated transcription factor (Foxp3) and cytokine (IL-10) at mRNA level was measured by real-time fluorescent quantitative polymerase chain reaction. Western blot and enzyme-linked immunosorbent assay ( ELISA) were used to detect the expression of Foxp3 and IL-10 at pro-tein level, respectively. Results Compared with the WT control group, the WT colitis group showed signif-icantly decreased proportions of Treg cells in PBMCs [(1. 85±0. 38)％ vs (3. 12±0. 69)％, P<0. 05], but increased proportions in MLNs [(11. 79±1. 18)％ vs (6. 24±1. 04)％, P<0. 05]. Compared with the WT mice with colitis, the TRAIL-/- mice with colitis had more severe colonic inflammation and significantly in-creased proportions of Treg cells in PBMCs [(3. 15±0. 64)％ vs (1. 85±0. 38)％, P<0. 05], but de-creased Treg cells in MLNs [(9. 80±0. 50)％ vs (11. 79±1. 18)％, P<0. 05]. Expression of Foxp3 and IL-10 at mRNA and protein levels in PBMCs of the WT mice with colitis was significantly lower than that in the WT control mice [ Foxp3 mRNA: 0. 48 ± 0. 21 vs 1. 06 ± 0. 31, IL-10 mRNA: 0. 23 ± 0. 07 vs 1. 22 ± 0. 38;Foxp3 protein:0. 68±0. 12 vs 1, IL-10 protein:(4. 91± 0. 72) pg/ml vs (21. 86±2. 40) pg/ml;all P<0. 05], while in MLNs, the expression of Foxp3 and IL-10 at mRNA and protein levels was significantly higher than that of the WT control group [Foxp3 mRNA:3. 71±0. 49 vs 1. 03±0. 15, IL-10 mRNA:11. 98 ±6.10 vs 1. 01±0. 31; Foxp3 protein: 1. 60±0. 03 vs 1, IL-10 protein: (1260. 00±18. 02) pg/ml vs (1184. 00±38. 62) pg/ml;all P<0. 05]. Compared with the WT mice with colitis, the TRAIL-/-mice with colitis showed significantly increased expression of Foxp3 and IL-10 at mRNA and protein levels [ Foxp3 mRNA:1. 80±0. 49 vs 0. 48±0. 21, IL-10 mRNA:1. 67±0. 99 vs 0. 23±0. 07;Foxp3 protein:1. 10±0. 01 vs 0. 68±0. 12, IL-10 protein:(31. 33± 25. 02) pg/ml vs (4. 58±3. 73) pg/ml; all P<0. 05], while de-creased expression in MLNs [ Foxp3 mRNA: 0. 49 ± 0. 21 vs 3. 71 ± 0. 49, IL-10 mRNA: 2. 80 ± 1. 82 vs 11. 98±6. 10; Foxp3 protein: 1. 21±0. 12 vs 1. 60±0. 03, IL-10 protein: (1158. 00±26. 48) pg/ml vs (1190. 00±37. 19) pg/ml;all P<0. 05]. Conclusions Knocking out the expression of TRAIL might af-fect the ratios of Treg cells in peripheral blood and MLNs, thereby aggravating the colitis in mice.

The Effect of Thymosin Alpha 1 on Severe Acute Pancreatitis in Rats / Estudios sobre el efecto de la timosina alfa 1 sobre la pancreatitis aguda grave en ratas

ABSTRACT Objective: To observe the effect of thymosin alpha l (Tα1) on severe acute pancreatitis (SAP) in rats. Methods: Twenty-four adult male Sprague-Dawley rats were randomly divided into three groups (eight in each group): control group (Group A), SAP group (Group B) and Tα1 treatment group (Group C). Animal models of SAP were made by retrograde injection of 5% sodium taurocholate into the biliopancreatic duct. Rats in Group C were treated with Tα1 (6 mg/kg) via intraperitoneal administration prior to SAP modelling. Eight rats in each group were sacrificed at 12 hours, respectively, after modelling. The serum levels of amylase, tumour necrosis factor-α (TNF-α), interleukin-lβ (IL-lβ and interleukin-6 (IL-6) were detected in each group. The pathological scores of the tissue in the pancreas head were observed by light microscopy. Results: The levels of serum amylase of Group B were 6378 ± 538 U/L, which were significantly higher than those (4587 ± 478 U/L) of Group C (p < 0.05). The levels of serum TNF-α of Group B were 360.32 ± 28.67 pg/mL, which were higher than those (269.99 ± 26.11 pg/mL) of Group C (p < 0.05). The levels of serum IL-lβ of Group B were 435.93 ± 36.00 pg/mL, which were higher than those (312.42 ± 17.89 pg/mL) of Group C (p < 0.05). The levels of serum IL-6 of Group B were 433.90 ± 28.36 pg/mL, which were higher than those (289.98 ± 23.00 pg/mL) of Group C (p < 0.05). The pancreatic pathological scores of Group B were 13.34 ± 2.19, which were higher than those (6.39 ± 1.86) of Group C (p < 0.05). Conclusion: Thymosin alpha 1 could decrease proinflammatory cytokines and reduce pancreas injury and had a protective effect in rats with SAP. This provides a new strategy for the clinical treatment of SAP.

RESUMEN Objetivo: Observar el efecto de la timosina alfa l (Tα1) sobre la pancreatitis aguda grave (PAG) en ratas. Métodos: Veinticuatro ratas Sprague-Dawley adultas machos fueron divididas aleatoriamente en tres grupos (ocho en cada grupo): grupo de control (grupo A), grupo de PAG (grupo B) y grupo de tratamiento con Tα1 (grupo C). Los modelos animales de PAG fueron creados mediante inyección retrógrada de taurocolato de sodio al 5% en el conducto biliopancreático. Las ratas del grupo C se trataron con Tα1 (6 mg/kg) via administración intraperitoneal antes del modelado de PAG. Las ocho ratas en cada grupo fueron sacrificadas a las 12 horas, respectivamente, después del modelado. Los niveles séricos de amilasa, factor-α de necrosis tumoral (TNF-α), interleucina-β (Il-β) e interleucina-6 (IL-6) fueron detectados en cada grupo. Las puntuaciones patológicas del tejido en la cabeza del páncreas fueron observadas mediante microscopía de luz. Resultados: Los niveles de amilasa sérica del grupo B fueron 6378 ± 538 U/L, y resultaron significativamente más altos (p < 0.05) que los niveles 4587 ± 478 U/L del grupo C. Los niveles séricos de TNF-α del grupo B fueron 360.32 ± 28.67 pg/mL, y resultaron ser más altos (p < 0.05) que los 269.99 ± 26.11 pg/mL del grupo C. Los niveles séricos de Il-β del grupo B fueron 435.93 ± 36.00 pg/mL, y fueron más altos (p < 0.05) que los 312.42 ± 17.89 pg/mL) del grupo C. Los niveles de suero IL-6 del grupo B fueron 433.90 ± 28.36 pg/mL, y resultaron ser más altos (p < 0.05) que los 289.98 ± 23.00 pg/mL del grupo C. Las puntuaciones patológicas pancreáticas del grupo B fueron 13.34 ± 2.19, y resultaron ser más altas (p < 0.05) que las puntuaciones 6.39 ± 1.86 del grupo C. Conclusión: La timosina alfa pudo disminuir las citoquinas proinflamatorias y reducir la lesión del páncreas, y tuvo un efecto protector en las ratas con PAG. Esto ofrece una nueva estrategia para el tratamiento clínico de PAG.

A fascinating story of the discovery & development of biologicals for use in clinical medicine

A young physician starting a fresh career in medicine in this millennium would hardly stop to think about the genesis of a particular biological drug that he/she will be prescribing for a patient evaluated in the morning outpatient department. For him/her, this is now routine, and the question of ‘Who’, ‘How’ and ‘When’ about these biologicals would be the last thing on their mind. However, for those who came to the medical profession in the 1950s, 1960s and 1970s, these targeted drugs are nothing short of ‘miracles’. It would be a fascinating story for the young doctor to learn about the long journey that the dedicated biomedical scientists of yesteryears took to reach the final destination of producing such wonder drugs. The story is much like an interesting novel, full of twists and turns, heart-breaking failures and glorious successes. The biologicals acting as ‘targeted therapy’ have not only changed the natural history of a large number of incurable/uncontrollable diseases but have also transformed the whole approach towards drug development. From the classical empirical process, there is now a complete shift towards understanding the disease pathobiology focusing on the dysregulated molecule(s), targeting them with greater precision and aiming for better results. Seminal advances in understanding the disease mechanism, development of remarkably effective new technologies, greater knowledge of the human genome and genetic medicine have all made it possible to reach the stage where artificially developed ‘targeted’ drugs are now therapeutically used in routine clinical medicine.

Correlation of transforming growth factor-b and tumour necrosis factor levels with left ventricular function in Chagas disease

BACKGROUND Transforming growth factor β1 (TGF-β1) and tumour necrosis factor (TNF) have been implicated in Chagas disease pathophysiology and may correlate with left ventricular (LV) function. OBJECTIVES We determined whether TGF-β1 and TNF serum levels correlate with LV systolic and diastolic functions and brain natriuretic peptide (BNP) serum levels in chronic Chagas disease. METHODS This cross-sectional study included 152 patients with Chagas disease (43% men; 57 ± 12 years old), classified as 53 patients with indeterminate form and 99 patients with cardiac form (stage A: 24, stage B: 25, stage C: 44, stage D: 6). TGF-β1, TNF, and BNP were determined by enzyme-linked immunosorbent assay ELISA. Echocardiogram was used to determine left atrial and LV diameters, as well as LV ejection fraction and diastolic function. FINDINGS TGF-b1 serum levels were lower in stages B, C, and D, while TNF serum levels were higher in stages C and D of the cardiac form. TGF-β1 presented a weak correlation with LV diastolic function and LV ejection fraction. TNF presented a weak correlation with left atrial and LV diameters and LV ejection fraction. CONCLUSIONS TNF is increased, while TGF-β1 is decreased in the cardiac form of chronic Chagas disease. TNF and TGF-β1 serum levels present a weak correlation with LV systolic and diastolic function in Chagas disease patients.

An experimental study on brain protective effect of Sini decoction on post-resuscitation syndrome / 中国中西医结合急救杂志

Objective To approach the brain protective effect and mechanism of Sini decoction on rats with cardiopulmonary resuscitation (CPR) syndrome.Methods Fifty Sprague-Dawley (SD) rats were divided into sham operation group (n = 10), CPR model group (n = 20) and Sini decoction treatment group (n = 20) by random number table. The rat models were established by trachea clipping to induce cardiac arrest, and after heart beat stopped for 5 minutes, CPR was carried out. In the Sini decoction group, Sini decoction 5 g/kg was given through a stomach tube, once per 24 hours, while in the sham and CPR model groups, the same amount of normal saline was given by the same method at the same time. Venous blood was collected before CPR and 6, 12, 24, 48 and 72 hours after CPR, and the levels of serum interleukin-6 (IL-6) and tumour necrosis factor-α (TNF-α) were determined by enzyme linked immunosorbent test (ELISA); after CPR for 72 hours, the rat brain tissue was obtained from all the groups, the content of caspase-3 in brain tissue was detected by immunohistochemistry method, and its protein expression caspase-3 was detected by Western Blot; the apoptosis situation of brain tissue was detected by terminal deoxynucleotidyl transferase-mediated duTP nick end labeling (TUNEL).Results With the prolongation of time, the levels of IL-6 and TNF-α in CPR model and Sini decoction groups showed a tendency firstly increased and then decreased, IL-6 reached its peak value after resuscitation for 24 hours, while TNF-α reached its peak value after resuscitation for 48 hours, and both IL-6 and TNF-α were decreased after resuscitation for 72 hours ; beginning from 6 hours after resuscitation, the levels of serum IL-6 (ng/L: 61.79±1.31, 62.49±1.42 vs. 21.48±0.79) and TNF-α (ng/L: 48.32±1.98, 25.32±1.96 vs. 18.34±2.45) in CPR model and Sini decoction treatment groups were all significantly higher than those in sham group, since 12 hours after resuscitation, the level of IL-6 was significantly lower in Sini decoction than that in CPR model group (ng/L: 70.41±2.21 vs. 88.32±1.59), and since 6 hours after resuscitation, TNF-α was obviously lower in Sini decoction group than that in CPR model group (ng/L: 25.32±1.96 vs. 48.32±1.98, allP < 0.05), both IL-6, TNF-α persisting to 72 hours after resuscitation, and their levels did not return to normal at the end of experiment in the two groups. After the end of resuscitation, the content and protein expression of caspase-3 and rate of cell apoptosis in the brain tissue in CPR model group were significantly higher than those in the sham group [caspase-3 content (A value,×103): 2.59±0.26 vs. 1.57±0.06, caspase-3 protein (gray value): 0.80±0.08 vs. 0.43±0.04, apoptosis rate: (2.01±0.08)% vs. (0.26±0.02)%, allP < 0.05], above indexes in the Sini decoction treatment group were significantly lower than those in the CPR model group [caspase-3 content (A value,×103): 1.89±0.08 vs. 2.59±0.26, caspase-3 protein (gray value): 0.57±0.02 vs. 0.80±0.08, apoptosis rate: (1.03±0.05)% vs. (2.01±0.08)%, allP < 0.05).Conclusion The Sini decoction has a protective effect on rats with post-resuscitation syndrome, and its mechanism is possibly realized by the inhibition of inflammatory factors and reduction of cell apoptosis.

An analysis of clinical efficacy of edaravone combined with β-aescin sodium for treatment of elderly patients with acute cerebral hemorrhage based on dynamic changes of vascular endothelial growth factor and inflammatory factor levels / 中国中西医结合急救杂志

Objective To investigate the clinical efficacy of edaravone combined with β-aescin sodium for treatment of elderly patients with acute cerebral hemorrhage,and the dynamic changes of tumor necrosis factor-α(TNF-α) and C-reactive protein (CRP),vascular endothelial growth factor (VEGF) levels in serum in such patients before and after treatment.Methods A total of 99 elderly patients with acute cerebral hemorrhage admitted to Henan Nanyang Second General Hospital from April 2014 to February 2016 were enrolled,and they were divided into control group (49 cases) and observation group (50 cases) according to the random digital table.Conventional treatment were given to both groups,patients in the observation group were added with edaravone 30 mg intravenous drip,2 times a day and β-aescin 20 mg intravenous drip,once daily.After treatment for consecutive 2 weeks,the clinical effect and neurological deficit score (NDS) were observed in the two groups;serum CRP levels were detected by immunonephelometry;TNF-αα and VEGF were detected by enzyme linked immunosorbent assay (ELISA).The changes of CRP,VEGF,TNF-α levels and incidence of adverse reactions before and after treatment were compared between the two groups.Results The levels of NDS,CRP,VEGF and TNF-α in both groups after treatment were significantly lower than those before treatment,and the degrees of decrease in treatment group were more significant than those in control group [NDS score:13.01 ± 1.37 vs.20.63 ± 1.68,CRP (mg/L):8.05 ± 3.97 vs.10.04 ± 4.17,VEGF (ng/L):97.25 ± 13.73 vs.116.43 ± 14.10,TNF-α (ng/L):8.15 ± 2.52 vs.11.54 ± 2.22,all P ＜ 0.05];the total effective rate in observation group was significantly higher than that in control group [92.00％ (46/50) vs.77.55％ (38/49),P ＜ 0.05].The difference of incidence of adverse reaction between observation group and control group was not statistically significant [4.00％ (2/50) vs.8.16％ (4/49),P ＞ 0.05].Conclusions Edaravone combined with β-aescin sodium for treatment of elderly patients with acute cerebral hemorrhage can effectively promote the recovery of neurological function,and its rolemight be related to the regulation of the levels of CRP,TNF-α and VEGF.

Relationship of Traditional Chinese Medical Syndromes of Type 2 Diabetic Nephropathy with Inflammatory Factors / 广州中医药大学学报

Objective To explore the relationship between traditional Chinese medicine(TCM) syndrome types of diabetic nephropathy(DN) patients and inflammatory factors, thus to supply evidence for syndrome differentiation, clinical treatment, illness evaluation and prognosis of DN patients. Methods The study was carried out in 120 cases of DN patients, and the patients were differentiated into TCM syndromes according to the clinical manifestations. Blood and urine samples were detected with enzyme-linked immunosorbent assay (ELISA). The correlation of syndrome types with blood and urine tumour necrosis factor-like weak inducer of apoptosis (TWEAK), interleukin-1β(IL-1β) and IL-10 was analyzed, and the detection results were compared to 30 healthy volunteers. Results (1) Compared with the healthy control, abnormal blood and urine TWEAK content and abnormal blood IL-1β were shown in DN patients at the stages of non-albuminuria, small-amount albuminuria, large-amount albuminuria, and renal insufficiency(P<0.05); blood IL-10 content was increased in DN patients without albuminuria(P<0.05).(2) DN patients with yin-deficiency and dryness-heat had higher blood and urine TWEAK contents than DN patients with other syndrome types (P<0.05). The blood TWEAK content was in decreasing sequence in the syndrome types of yin-deficiency and dryness-heat, Qi-yin deficiency, spleen-kidney Qi deficiency, yin-yang deficiency; the urine TWEAK content was in decreasing sequence in the syndrome types of yin-deficiency and dryness-heat, Qi-yin deficiency, spleen-kidney Qi deficiency, yin-yang deficiency(P<0.05). DN patients with damp-heat syndrome had the highest blood IL-1βcontent (P < 0.05), and yin-deficiency and dryness-heat had the lowest IL-1β content (P < 0.05). DN patients with yin-deficiency and dryness-heat had higher blood IL-10 content (P<0.05), and the blood IL-10 content was in decreasing sequence in the syndrome types of yin-deficiency and dryness-heat, Qi-yin deficiency, spleen-kidney Qi deficiency, yin-yang deficiency. Conclusion TCM syndrome types of DN patients are correlated with TWEAK, IL-1β and IL-10, and the results will supply evidence for syndrome differentiation, clinical treatment, illness evaluation and prognosis of DN patients .