Effect of siRNA interference with C-C chemokine ligand 5 on biological characteristics of laryngeal carcinoma cells / 中国生物制品学杂志

@#Objective To investigate the expression of C-C chemokine ligand 5(CCL5) in head and neck squamous cell carcinoma(HNSCC),and explore the effect of CCL5 on the biological characteristics of laryngeal carcinoma cells.Methods Gene Expression Profiling Interactive Analysis(GEPIA) database was used to investigate the expression of CCL5 in HNSCC.The laryngeal carcinoma cells TU177 were transfected with siRNA(siRNA group),and the control(NC) group was set up.The cell proliferation,migration,cycle and apoptosis of each group were detected by CCK8 assay,cell scratch test and flow cytometry respectively.RT-PCR and Western blot were used to detect the knock-down efficiency of CCL5 and the mRNA transcription and protein expression of multidrug resistance protein 2(MRP2) and bcl-2-associated x protein(Bax).Results The expression of CCL5 in HNSCC was higher than that in normal tissues(P <0.05).Compared with NC group,siRNA showed higher knock-down efficiency(t=12.898 and 22.656 respectively,each P <0.01);siRNA interference with CCL5 inhibited the proliferation and migration of laryngeal carcinoma cells,and promoted the late apoptosis of laryngeal carcinoma cells and the expression of apoptosis protein Bax(t=2.600～11.667,each P <0.05).Conclusion CCL5 was highly expressed in HNSCC,while siRNA interference with CCL5 inhibited the proliferation,migration and promoted apoptosis of laryngeal carcinoma cells TU177 by up-regulating the expression of Bax,which laid a foundation of the possibility of CCL5 as a new target for the treatment of laryngeal carcinoma.

Regulation of Baicalin on Growth of Extranodal NK/T Cell Lymphoma Cells through FOXO3/CCL22 Signaling Pathway / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect of baicalin on the growth of extranodal NK/T cell lymphoma (ENKTCL) cells and its related mechanism.@*METHODS@#Normal NK cells and human ENKTCL cells lines SNK-6 and YTS were cultured, then SNK-6 and YTS cells were treated with 5, 10, 20 μmol/L baicalin and set control. Cell proliferation and apoptosis was detected by Edu method and FCM method, respectively, and expressions of BCL-2, Bax, FOXO3 and CCL22 proteins were detected by Western blot. Interference plasmids were designed and synthesized. FOXO3 siRNA interference plasmids and CCL22 pcDNA overexpression plasmids were transfected with PEI transfection reagent. Furthermore, animal models were established for validation.@*RESULTS@#In control group and 5, 10, 20 μmol/L baicalin group, the proliferation rate of SNK-6 cells was (56.17±2.96)%, (51.92±4.63)%, (36.42±1.58)%, and (14.60±2.81)%, respectively, while that of YTS cells was (58.85±2.98)%, (51.38±1.32)%, (34.75±1.09)%, and (15.45±1.10)%, respectively. In control group and 5, 10, 20 μmol/L baicalin group, the apoptosis rate of SNK-6 cells was (5.93±0.74)%, (11.78±0.34)%, (28.46±0.44)%, and (32.40±0.37)%, respectively, while that of YTS cells was (7.93±0.69)%, (16.29±1.35)%, (33.91±1.56)%, and (36.27±1.06)%, respectively. Compared with control group, the expression of BCL-2 protein both in SNK-6 and YTS cells decreased significantly (P<0.001), and the expression of Bax protein increased in SNK-6 cells only when the concentration of baicalin was 20 μmol/L (P<0.001), while that in YTS cells increased in all three concentrations(5, 10, 20 μmol/L) of baicalin (P<0.001). The expression of FOXO3 protein decreased while CCL22 protein increased in ENKTCL cell lines compared with human NK cells (P<0.001), but the expression of FOXO3 protein increased (P<0.01) and CCL22 protein decreased after baicalin treatment (P<0.001). Animal experiments showed that baicalin treatment could inhibit tumor growth. The expression of CCL22 protein in ENKTCL tissue of nude mice treated with baicalin decreased compared with control group (P<0.01), while the FOXO3 protein increased (P<0.05). In addition, FOXO3 silencing resulted in the decrease of FOXO3 protein expression and increase of CCL22 protein expression (P<0.01, P<0.001).@*CONCLUSION@#Baicalin can inhibit proliferation and promote apoptosis of ENKTCL cell lines SNK-6 and YTS, up-regulate the expression of Bax protein, down-regulate the expression of BCL-2 protein, and down-regulate the expression of CCL22 protein mediated by FOXO3. Animal experiment shown that the baicalin can inhibit tumor growth. Baicalin can inhibit the growth and induce apoptosis of ENKTCL cells through FOXO3/CCL22 signaling pathway.

Catechin interferes with SCD1 expression and prevents liver fibrosis in mice / 中国药理学通报

Aim To investigate whether catechin can play against CCl

Renin angiotensin system molecules and chemokine (C-C motif) ligand 2 (CCL2) in chronic kidney disease patients / Moléculas do sistema renina-angiotensina e ligante 2 de quimiocina com motivo C-C (CCL2) em pacientes com doença renal crônica

Abstract Introduction: Studies have shown that the renin angiotensin aldosterone system (RAAS) and inflammation are related to kidney injury progression. The aim of this study was to evaluate RAAS molecules and chemokine (C-C motif) ligand 2 (CCL2) in 82 patients with chronic kidney disease (CKD). Methods: Patients were divided into two groups: patients diagnosed with CKD and patients without a CKD diagnosis. Glomerular filtration rate (GFR) and albumin/creatinine ratio (ACR) were determined, as well as plasma levels of angiotensin-(1-7) [Ang-(1-7)], angiotensin-converting enzyme (ACE)1, ACE2, and plasma and urinary levels of CCL2. Results: CCL2 plasma levels were significantly higher in patients with CKD compared to the control group. Patients with lower GFR had higher plasma levels of ACE2 and CCL2 and lower ratio ACE1/ACE2. Patients with higher ACR values had higher ACE1 plasma levels. Conclusion: Patients with CKD showed greater activity of both RAAS axes, the classic and alternative, and higher plasma levels of CCL2. Therefore, plasma levels of RAAS molecules and CCL2 seem to be promising prognostic markers and even therapeutic targets for CKD.

Resumo Introdução: Estudos têm mostrado que o sistema renina angiotensina aldosterona (SRAA) e a inflamação estão relacionados à progressão da lesão renal. O objetivo deste estudo foi avaliar moléculas do SRAA e o Ligante 2 de Quimiocina com Motivo C-C (CCL2) em 82 pacientes com doença renal crônica (DRC). Métodos: Os pacientes foram divididos em dois grupos: pacientes diagnosticados com DRC e pacientes sem diagnóstico de DRC. Foram determinadas a taxa de filtração glomerular (TFG) e a relação albumina/creatinina (RAC), assim como os níveis plasmáticos de angiotensina-(1-7) [Ang-(1-7)], enzima conversora de angiotensina (ECA)1, ECA2 e níveis plasmáticos e urinários de CCL2. Resultados: Os níveis plasmáticos de CCL2 foram significativamente mais altos em pacientes com DRC em comparação com o grupo controle. Pacientes com TFG mais baixa apresentaram níveis plasmáticos mais elevados de ECA2 e CCL2 e menor relação ECA1/ECA2. Pacientes com valores de RAC mais altos apresentaram níveis plasmáticos de ECA1 mais elevados. Conclusão: Pacientes com DRC mostraram maior atividade de ambos os eixos do SRAA, o clássico e o alternativo, e níveis plasmáticos mais altos de CCL2. Portanto, os níveis plasmáticos de moléculas do SRAA e CCL2 parecem ser marcadores prognósticos promissores e até mesmo alvos terapêuticos para a DRC.

Relationship between CCL21 and TREM2/DAP12 signaling pathways in spinal dorsal horn in remifentanil-induced hyperalgesia in mice with incisional pain / 中华麻醉学杂志

Objective:To evaluate the relationship between CCL21 and triggering receptor expressed on myeloid cells 2 (TREM2)/DNAX-activating protein of 12 kDa (DAP12) signaling pathways in the spinal dorsal horn in remifentanil-induced hyperalgesia in mice with incisional pain.Methods:Thirty-two SPF healthy male C57BL/6J mice, weighing 18-22 g, aged 8-10 weeks, were divided into 4 groups ( n=8 each) using a random number table method: control group (group C), CCL21 neutralizing antibody group (group anti-CCL21), remifentanil + incisional pain group (group R+ I), and CCL21 neutralizing antibody + remifentanil + incisional pain group (group anti-CCL21+ R+ I).A CCL21 neutralizing antibody 0.3 μg (diluted to 10 μl in normal saline) was intrathecally injected in anti-CCL21 and anti-CCL21+ R+ I groups twice a day.Normal saline 10 μl was intrathecally injected at the same time point twice a day in C and R+ I groups.Fifteen min after intrathecal injection, normal saline 0.1 ml was injected via the caudal vein for 4 consecutive times at an interval of 15 min in C and anti-CCL21 groups, and remifentanil 10 μg/kg (diluted to 0.1 ml in normal saline) was injected via the caudal vein for 4 consecutive times at an interval of 15 min in R+ I and anti-CCL21+ R+ I groups.The tail-flick latency (TFL) and mechanical paw withdrawal threshold (MWT) were measured at 24 h before remifentanil or normal saline injection (T 0) and 3, 6, 24 and 48 h after stopping injection of remifentanil or normal saline (T 1-4).The mice were sacrificed after the last measurement of pain threshold, and L 4-6 segments of the spinal cord were removed for determination of the expression of TREM2 and DAP12 protein and mRNA (by Western blot or quantitative real-time polymerase chain reaction). Results:Compared with group C, TFL was significantly shortened and MWT was decreased at T 1-4, and the expression of TREM2 and DAP12 protein and mRNA was up-regulated in group R+ I and R+ I+ anti-CCL21 ( P<0.05), and no significant change was found in the parameters mentioned above in group anti-CCL21 ( P>0.05).Compared with group R+ I, TFL was significantly prolonged and MWT was increased at T 1-4, and the expression of TREM2 and DAP12 protein and mRNA was down-regulated in group anti-CCL21+ R+ I ( P<0.05). Conclusions:CCL21 is involved in remifentanil-induced hyperalgesia by activating TREM2/DAP12 signaling pathways in the spinal dorsal horn of mice with incisional pain.

The postoperative cognitive dysfunction induced by central inflammation with possible involvement of the gut-brain axis

Abstract Background: Postoperative cognitive dysfunction is widely recognized as severe postoperative central nervous dysfunction and has a significant impact on the 'patient's physical and mental health. Methods: Postoperative models of tibial fracture in aged rats were established, including the control group, model group, CCL11 protein injection group, and saline injection group. Morris water maze test was used to detect the behavioral characteristics of rats. Enzyme-Linked Immunosorbent Assay was used or determine the content of CCL11 and CXCL10. Immunofluorescence staining was used to detect the distribution of CD14+CD163+macro-phages in colon tissues and CD11b+CCR3+microglia cells in hippocampal tissues. Western blot analyzed NOX1 and STAT3 expression in hippocampus tissues. Results: Water maze test results confirmed severe cognitive impairment in CCL11 rats. The content of CCL11 and CXCL10 in the CCL11 group was much higher than that of the model group. The distribution of macrophage and microglia cells in the CCL11 model group was greater than that in the model group and the saline group. The expression of NOX1 and STAT3 in the CCL11 group was higher compared with the model group. Conclusion: Abnormal macrophage function and excessive CCL11 secretion were observed in the rats with lower limb fractures after surgery. Postoperative central inflammation in rats with lower limb fracture induced postoperative cognitive dysfunction through the gut-brain axis molecular mechanism.

Progress on the mechanism of chemokine CCL2 in the malignant progression of glioma / 肿瘤研究与临床

Glioma is the most common primary intracranial tumor. At present, the conventional treatment methods have limited effect and cannot significantly prolong the survival time of patients. Chemokine CCL2 is the most important member of the CC chemokine family, which can regulate glioma angiogenesis, immunosuppression, progression and invasion, and resistance to apoptosis. This article reviews the potential mechanism of CCL2 promoting the malignant progression of glioma, in order to provide new ideas and methods for the targeted therapy of glioma.

Correlations between pruritus and CC chemokine ligand 17 in patients with mycosis fungoides / 中华皮肤科杂志

Objective:To investigate molecules involved in the occurrence of pruritus in patients with mycosis fungoides (MF) .Methods:Totally, 522 patients with MF were enrolled from Peking University First Hospital from October 2009 to August 2021, and the incidence of pruritus was calculated. The patients were grouped according to whether they suffered from pruritus or not. RNA sequencing data on biopsied skin lesions of 49 patients were analyzed to identify differentially expressed genes between patients with pruritus and those without; enzyme-linked immunosorbent assay and immunohistochemical techniques were performed to determine the protein expression of CC chemokine ligand 17 (CCL17) in serum samples from 88 MF patients, and in tissue samples from 81 MF patients, respectively; flow cytometry was conducted to detect markers for T lymphocyte activation and differentiation in peripheral blood samples from 46 MF patients to identify peripheral blood lymphocyte subsets associated with pruritus. Statistical analysis was carried out by using chi-square test, Mann-Whitney U test, and Spearman correlation analysis. Results:Among the 522 patients with MF, 305 were males and 217 were females; 347 were diagnosed with early-stage MF, and 175 with advanced MF. The incidence of pruritus was 67.2% (351/522) in the patients with MF, and significantly higher in the patients with advanced MF (81.7%, 143/175) than in those with early-stage MF (59.9%, 208/347; χ2 = 25.03, P < 0.001) . RNA sequencing showed that CCL17 mRNA expression was significantly higher in the MF patients with pruritus than in those without (fold change = 10.09, P < 0.001) . The serum CCL17 concentration was significantly elevated in the patients with pruritus (1 017.05[377.12, 4 831.80] pg/ml) compared with those without (361.66 [180.47, 500.08] pg/ml; Z = -4.57, P < 0.001) , and correlated with pruritus scores ( r = 0.57, P = 0.010) . In both early and advanced stages of MF, the serum CCL17 concentration was significantly higher in the patients with pruritus than in those without ( Z = -3.68, P < 0.001; Z = -2.54, P = 0.011, respectively) . Immunohistochemical staining revealed that there was no significant difference in the relative quantification value of CCL17 between the patients with pruritus and those without ( Z = -1.84, P = 0.066) . The percentage of CD3 +CD4 +CD26 -CCR4 + malignant T cells significantly increased in the MF patients with pruritus than in those without ( Z = -2.03, P = 0.043) , and was positively correlated with serum CCL17 concentrations ( r = 0.49, P < 0.001) . Conclusions:Both CCL17 mRNA expression in lesional tissues and serum CCL17 concentrations increased in MF patients with pruritus, and CCL17 was associated with the occurrence of pruritus. CCL17 may be involved in the occurrence of pruritus through the recruitment of CD3 +CD4 +CD26 -CCR4 + malignant T cells.

Comparison of two methods for establishing non-alcoholic fatty liver disease model in mice / 中国药理学通报

Aim To compare the effects of two different methods of establishing non-alcoholic fatty liver disease ( NAFLD) model, and to explore a more efficient method for establishing NAFLD model that conforms to the characteristics of human disease.Methods C57BL/6J mice were randomly divided into NI) group, WD group, WD combined with CC14 group.'Hie NI) group was fed a normal maintenance diet, and WD group was fed WD.On the basis of WD feed, mice in WD + CC14 group were intraperitoneally injected with CC14 oil solution.'Hie mice were sacrificed on 6, 11 and 16 weeks after modeling.HE staining and oil red 0 staining were performed to observe the pathological changes of liver.The serum levels of ALT, AST, TG and T- CHO were detected by automatic biochemical analyzer, and the levels of IL-lp, IL-6 and TNF-a in liver homogenate were detected by ELISA.The protein expression of FAS and a-SMA was detected by Western blot.Results As the development of model, pathological results showed that NAFLD model was success-fully established by these two methods.At the same time point of modeling, compared with WD group, the liver pathology of WD + CC14 group was more serious, liver steatosis appeared since 6th week.The serum ALT, AST levels and the contents of TG and T-CHO significantly increased.Meanwhile, the levels of inflammatory cytokines obviously increased in the liver, the expression of fibrosis-associated protein a-SMA increased, and the model could progress to the stage of NASH on 16th week.The course of NAFLD in the WD group progressed slowly, and steatosis appeared on 1 1 th week, and it was further aggravated till 16th week.The pro-tein level of FAS was significantly higher than that in WD + CC14 group, and no obvious inflammatory cell infiltration was observed.Conclusions WD feed combined with CC14 to establish NAFLD model takes shorter time and exerts better effect than feeding WD a- lone.It can progress to NASH on 16th week, which can be used as an ideal method to establish NAFLD model.

Protective effects of bicistronic DNA vaccines of herpes simplex virus type 2 glycoprotein D and adjuvant CCL28 on mice / 中华微生物学和免疫学杂志

Objective:To study the protective effects of bicistronic DNA vaccines carrying herpes simplex virus type 2 glycoprotein D (HSV-2 gD) and adjuvant CCL28 sequences that were connected by internal ribosome entry site (IRES) sequence in mouse model.Methods:The recombinant DNA vaccines, pgD-IRES-CCL28 and pCCL28-IRES-gD, encoding HSV-2 gD and adjuvant CCL28 were constructed with IRES sequence. After verified by sequencing, they were intramuscularly injected twice into BALB/c mice. Serum samples and vaginal lavage fluids were collected regularly. Splenocytes, mesenteric lymph node cells and rectal mucosa tissues were separated and collected. The titers of antigen-specific antibodies in immunized mice were analyzed with end-point ELISA. In vitro neutralization assay was used to measure neutralizing antibody titers in serum and vaginal lavage fluid after vaccination and virus challenge. CCL28-responsive immune cells in splenocytes, mesenteric lymph node cells and rectal tissues were detected by chemotaxis experiment and immunohistochemical staining. The protective effects of the bicistronic DNA vaccines were evaluated by fluorescent quantitative PCR, weighing and disease severity assessment. Humoral and cellular immune responses induced by the bicistronic DNA vaccines and their efficacy in immunoprotection were analyzed by comparing with pgD+ pCCL28 group. Results:IgG titers in serum samples and IgA antibody titers in vaginal lavage fluids of mice immunized with pCCL28-IRES-gD were similar to those in pgD+ pCCL28 group. The neutralizing ability of antibodies, the number of rectal mucosal IgA+ plasma cells and CCL28-responsive immune cells in mucosal tissues were increased in pCCL28-IRES-gD group. Serum neutralizing antibodies were not produced immediately in the mice challenged with HSV-2, but no weight loss, disease symptoms or death was observed. However, pgD+ pcDNA3.1 and pgD-IRES-CCL28 were ineffective against HSV-2 infection in mice.Conclusions:The recombinant bicistronic DNA vaccine of pCCL28-IRES-gD could induce stronger mucosal immune response in mice and provide better protective effects.

Protective effects of fusion recombinant DNA vaccine of herpes simplex virus type 2 glycoprotein D and adjuvant CCL19 on mice / 中华微生物学和免疫学杂志

Objective:To compare the immune responses to simply mixed and fused recombinant DNA vaccines of herpes simplex virus type 2 glycoprotein D (HSV-2 gD) and molecular adjuvant CCL19 in mice and to evaluate the protective effects.Methods:Gene recombination technology was used to construct recombinant DNA vaccines expressing HSV-2 gD and CCL19 alone or fused together. After verification by sequencing, Western blot and ELISA, BALB/c mice were immunized twice by intramuscular injection. Serum samples and vaginal lavage fluids were collected regularly after immunization. Splenocytes, mesenteric lymph node cells and rectal tissues were collected after immunization. Differences in humoral and cellular immune responses to the two forms of vaccines and their protective effects in mice were analyzed using end-point ELISA, in vitro neutralization assay, immunohistochemical staining, chemotaxis assay, vaginal virus challenge, fluorescence quantitative PCR, weighing and disease severity assessment. Results:The fused recombinant pgD-IZ-CCL19 plasmid could express gD protein and CCL19 protein in vitro, but the level of expressed CCL19 protein by pCCL19-IZ-gD plasmid was less than that by pgD-IZ-CCL19. The mice immunized with pgD-IZ-CCL19 showed higher levels of IgG in sera and IgA in vaginal lavage fluids ( P<0.01) and stronger neutralization ability than the mice vaccinated with pgD+ pCCL19. Compared with other groups, more lymphocytes were recruited in the rectal mucosa, the spleen and mesenteric lymph nodes of mice immunized with pgD-IZ-CCL19. Weight loss or disease symptoms were not observed in the pgD-IZ-CCL19 group after virus challenge. In addition, the positive rate of HSV-2 in vaginal mucosa and the mortality rate in the pgD-IZ-CCL19 group were the lowest. However, pCCL19-IZ-gD turned out ineffective in preventing HSV-2 infection. Conclusions:The fused recombinant DNA vaccine pgD-IZ-CCL19 could induce stronger immune responses in mice and provide better protective effects, which was superior to the simply mixed DNA vaccine.

Effect of Shufeng Tongluo Prescription on Eotaxin and CCR3 Protein Expression and ERK Phosphorylation Level of Asthma Mice / 中国实验方剂学杂志

ObjectiveTo preliminarily explore the mechanism of Shufeng Tongluo prescription （SFTLP） in inhibiting airway inflammation in asthma mice by affecting the expression levels of eotaxin in the serum， CC type chemokine receptor 3 （CCR3）， and extracellular signal-regulated kinase （ERK） phosphorylation in lung tissues. MethodSeventy C57BL/6 mice were randomly divided into a blank group， a model group， low-， medium-， and high-dose SFTLP groups （7.75， 15.5， 30 g·kg-1）， a pertussis toxin （PTX） group， a CCR3 inhibitor （SB328437） group， a phosphoinositide 3-kinase （PI3K） inhibitor （LY294002） group， a p38 protein kinase antagonist inhibitor （SB203580） group， and an ERK inhibitor （PD98059） group. The asthma model was induced in mice by intraperitoneal injection of ovalbumin （OVA） and aluminum hydroxide ［Al（OH）3］ combined with OVA atomization （0.2 mL for all）. After modeling， hematoxylin-eosin staining （HE staining） was used to observe the inflammatory infiltration of lung tissues in mice. Enzyme-linked immunosorbent assay （ELISA） was used to detect the serum levels of eotaxin ［CC chemokine ligand （CCL） 11 and CCL24） in each group. Western blot was used to detect the levels of ERK phosphorylation and CCR3 in lung tissues. ResultCompared with the blank group， the model group showed obvious bronchial constriction， lumen stenosis， damaged alveolar structure， massive inflammatory cell infiltration in lung tissues， mucous plug in the bronchus， edema in the submucosal tissues of the trachea， increased folds， increased serum levels of CCL11 and CCL24 （P<0.01）， and increased expression of CCR3 protein in lung tissues （P<0.05）. The ERK levels in lung tissues of the model group and the PTX group increased （P<0.05）. The level of p-ERK in lung tissues of the model group and the low-dose SFTLP group increased （P<0.05）. As revealed by pathological results， compared with the model group， the high-dose SFTLP group showed relieved lung lesions. The high-dose SFTLP group and the SB328437 group showed reduced CCL11 content （P<0.05）. The low- and high-dose SFTLP group， the PTX group， the SB203580 group， the PD98059 group， and the SB328437 group showed decreased CCR3 protein expression in lung tissues （P<0.05）. The high-dose SFTLP group and the PD98059 group showed reduced p-ERK level （P<0.05）. The PD98059 group showed reduced ERK level （P<0.05）. ConclusionSFTLP can inhibit airway inflammation in asthma， and the mechanism may be related to the inhibition of eosinophil activation by down-regulating CCR3 and CCL11 expression and ERK phosphorylation.

Effect of Acute Liver Injury on Pharmacokinetics and Tissue Distribution of Baicalin Magnesium in Rats / 中国实验方剂学杂志

ObjectiveTo establish a high performance liquid chromatography （HPLC） for simultaneous determination of baicalin magnesium and baicalein in rat plasma and tissues， and to investigate the effect of acute liver injury on pharmacokinetics and tissue distribution of baicalin magnesium in rats. MethodAcute liver injury rat model was induced by carbon tetrachloride （CCl4）. Normal rats and acute liver injury model rats were given an equal dose （287.31 mg·kg-1） of baicalin magnesium aqueous solution by intragastric administration， the orbital blood was collected at different time points， and HPLC was used to simultaneously determine the concentrations of baicalin magnesium and baicalein in rat plasma at each time point， the concentration-time curves were drawn， the pharmacokinetic parameters were calculated with DAS 3.0， and SPSS 23.0 was used for statistical analysis. After oral administration of baicalin magnesium aqueous solution， HPLC was used to simultaneously determine the contents of baicalin magnesium and baicalein in rat liver， lung， kidney， stomach， brain and small intestine at different time points， the mobile phase was 0.1% phosphoric acid aqueous solution-methanol， and the detection wavelength was 278 nm. ResultIn the acute liver injury model group， the peak concentration （Cmax） of baicalin magnesium was 0.58 times that of the normal group， the area under concentration-time curve （AUC0-t） was 0.5 times that of the normal group （P<0.05）， the apparent volume of distribution （Vd） was 2.3 times that of the normal group （P<0.05）， and baicalein is almost undetectable in plasma. The content of baicalin magnesium in liver， stomach and brain of the acute liver injury model group was higher than that of the normal group at each time point， while the content of baicalin magnesium in the samples of lung at 8 h， kidney at 8 h and 12 h， and small intestine at 0.333 h was lower than that of the normal group. The content of baicalein in lung， stomach and small intestine of the model group was higher than that of the normal group at each time point， while the content of baicalein in the tissue samples of liver at 6， 8 h and kidney at 0.333， 4， 6 h was lower than that in the normal group， and baicalein could hardly be detected in the brain. ConclusionAfter intragastric administration of the same dose of baicalin magnesium aqueous solution， acute liver injury induced by CCl4 can affect the pharmacokinetics and tissue distribution characteristics of baicalin magnesium in rats， and there is biotransformation of baicalin magnesium and baicalein in liver， lung， kidney， stomach and small intestine.

Mechanism of Zhizi Prescription in Protection of CCl4-induced Acute and Subacute Liver Injury in Mice / 中国实验方剂学杂志

ObjectiveTo investigate the protective effect of Zhizi prescription （ZZP） on carbon tetrachloride （CCl4）-induced acute and subacute liver injury and its mechanism. MethodAcute and subacute liver injury animal models were induced. C57 mice were randomly divided into a normal group， model group， obeccholic acid group， ZZP high-dose （0.5 g·kg-1） group， and ZZP low-dose （0.25 g·kg-1） group. According to the experiment design， the serum and liver tissue of mice were collected after the last administration. Hematoxylin-eosin （HE） and Sirius staining was used to observe the liver pathological changes. Serum alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， total bilirubin （TBIL）， liver homogenate hydroxyproline （Hyp）， malondialdehyde （MDA）， and superoxide dismutase （SOD） levels were determined by kit. The levels of tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6） in the liver tissue were determined by enzyme-linked immunosorbent assay （ELISA）. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect the mRNA expressions of collagen 1A1 （Col1a1）， collagen 3A1 （Col3a1）， fibronectin （FN）， transforming growth factor β receptor Ⅱ （Tgfbr2） and α-smooth muscle actin （α-SMA） in the liver tissue. ResultIn terms of the acute liver injury， as compared with the normal group， the levels of ALT， AST， TBIL and MDA in the model group were significantly increased （P<0.01）， while the activity of liver SOD was significantly decreased （P<0.01）. Compared with model group， the ZZP high-dose and low-dose groups both significantly reduced the degree of liver cell injury， and protected the acute liver injury induced by CCl4. The ZZP high-dose group had a better effect than the ZZP low-dose group. In terms of the subacute liver injury， the levels of ALT， AST， MDA，TNF-α and IL-6 in the model group were significantly increased （P<0.01）， while the activity of liver SOD was significantly decreased （P<0.01）. As compared with the model group， liver Hyp content in the ZZP high-dose and low-dose groups was significantly decreased （P<0.01）， and the collagen deposition in liver of both groups was significantly reduced. The ZZP high-dose group also significantly down-regulated the mRNA expressions of α-SMA， Col1a1， Col3a1， FN， and Tgfbr2 in the liver of mice （P<0.05， P<0.01）. ConclusionZZP effectively protects the acute and subacute liver injury induced by CCl4， and the protective effect is proportional to its concentration. The mechanism may be related to the increase of the activity of antioxidant enzymes in the liver tissue， the decrease of the level of lipid peroxidation， and the inhibition of inflammatory response， thus reducing collagen deposition and improving early liver fibrosis.

Mechanism of Zhizi Prescription in Protection of CCl4-induced Acute and Subacute Liver Injury in Mice / 中国实验方剂学杂志

ObjectiveTo investigate the protective effect of Zhizi prescription （ZZP） on carbon tetrachloride （CCl4）-induced acute and subacute liver injury and its mechanism. MethodAcute and subacute liver injury animal models were induced. C57 mice were randomly divided into a normal group， model group， obeccholic acid group， ZZP high-dose （0.5 g·kg-1） group， and ZZP low-dose （0.25 g·kg-1） group. According to the experiment design， the serum and liver tissue of mice were collected after the last administration. Hematoxylin-eosin （HE） and Sirius staining was used to observe the liver pathological changes. Serum alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， total bilirubin （TBIL）， liver homogenate hydroxyproline （Hyp）， malondialdehyde （MDA）， and superoxide dismutase （SOD） levels were determined by kit. The levels of tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6） in the liver tissue were determined by enzyme-linked immunosorbent assay （ELISA）. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect the mRNA expressions of collagen 1A1 （Col1a1）， collagen 3A1 （Col3a1）， fibronectin （FN）， transforming growth factor β receptor Ⅱ （Tgfbr2） and α-smooth muscle actin （α-SMA） in the liver tissue. ResultIn terms of the acute liver injury， as compared with the normal group， the levels of ALT， AST， TBIL and MDA in the model group were significantly increased （P<0.01）， while the activity of liver SOD was significantly decreased （P<0.01）. Compared with model group， the ZZP high-dose and low-dose groups both significantly reduced the degree of liver cell injury， and protected the acute liver injury induced by CCl4. The ZZP high-dose group had a better effect than the ZZP low-dose group. In terms of the subacute liver injury， the levels of ALT， AST， MDA，TNF-α and IL-6 in the model group were significantly increased （P<0.01）， while the activity of liver SOD was significantly decreased （P<0.01）. As compared with the model group， liver Hyp content in the ZZP high-dose and low-dose groups was significantly decreased （P<0.01）， and the collagen deposition in liver of both groups was significantly reduced. The ZZP high-dose group also significantly down-regulated the mRNA expressions of α-SMA， Col1a1， Col3a1， FN， and Tgfbr2 in the liver of mice （P<0.05， P<0.01）. ConclusionZZP effectively protects the acute and subacute liver injury induced by CCl4， and the protective effect is proportional to its concentration. The mechanism may be related to the increase of the activity of antioxidant enzymes in the liver tissue， the decrease of the level of lipid peroxidation， and the inhibition of inflammatory response， thus reducing collagen deposition and improving early liver fibrosis.

CA-125 and CCL2 may indicate inflammation in peritoneal dialysis patients / CA-125 e CCL2 podem indicar inflamação em pacientes em diálise peritoneal

Abstract Introduction: Progressive structural changes in the peritoneal membrane occur over the course of treatment in peritoneal dialysis (PD), resulting in an increase in cytokines such as CCL2 and structural changes in peritoneal membrane triggering an increase in CA-125 in dialysate, which reflects a probable local inflammatory process, with possible loss of mesothelial cells. Thus, the current study aimed to evaluate the association between plasma and CCL2 and CA-125 dialysate levels in patients undergoing PD. Methods: Cross-sectional study was conducted with 41 patients undergoing PD. The assessments of CA-125 and CCL2 levels were performed using a capture ELISA. Correlations were estimated using Spearman's correlation and the investigation of the association between the explanatory variables (CCL2) and response variable (CA-125) was done for crude ratio of arithmetic means and adjusted utilizing generalized linear models. Results: A moderate positive correlation was observed between the levels of CA-125 and CCL2 in the dialysate (rho = 0.696). A statistically significant association was found between the levels in the CCL2 and CA-125 dialysate (RoM=1.31; CI = 1.20-1.43), which remained after adjustment for age (RoM = 1.31; CI=1.19-1.44) and for time in months of PD (RoM=1.34, CI=1.22-1.48). Conclusion: The association of CA-125 levels with CCL2 in the dialysate may indicate that the local inflammatory process leads to temporary or definitive changes in peritoneal membrane. A better understanding of this pathogenesis could contribute to the discovery of new inflammatory biomarkers.

Resumo Introdução: Alterações estruturais progressivas na membrana peritoneal ocorrem no decorrer do tratamento em diálise peritoneal (DP), resultando em um aumento de citocinas como CCL2 e alterações estruturais na membrana peritoneal desencadeando um aumento de CA-125 no dialisato, o que reflete um provável processo inflamatório local, com possível perda de células mesoteliais. Assim, o presente estudo teve como objetivo avaliar a associação entre CCL2 e CA-125 no plasma e no dialisato de pacientes submetidos à DP. Métodos: Foi realizado um estudo transversal com 41 pacientes submetidos à DP. As avaliações dos níveis de CA-125 e CCL2 foram realizadas utilizando ELISA de captura. As correlações foram estimadas usando a correlação de Spearman, e a investigação da associação entre as variáveis explicativas (CCL2) e a variável resposta (CA-125) foi feita pela razão bruta das médias aritméticas e ajustada utilizando modelos lineares generalizados. Resultados: Foi observada uma correlação positiva moderada entre os níveis de CA-125 e CCL2 no dialisato (rho = 0,696). Foi encontrada uma associação estatisticamente significativa entre os níveis no dialisato de CCL2 e CA-125 (RoM=1,31; IC = 1,20-1,43), que permaneceu após ajuste por idade (RoM = 1,31; IC=1,19-1,44) e pelo tempo de DP em meses (RoM=1,34, IC=1,22-1,48). Conclusão: A associação dos níveis de CA-125 com CCL2 no dialisato pode indicar que o processo inflamatório local leva a alterações temporárias ou definitivas na membrana peritoneal. Uma melhor compreensão desta patogênese pode contribuir para a descoberta de novos biomarcadores inflamatórios.

Genes implicados en la gravedad de la infección por SARS-cov- 2

Resumen El COVID-19 es una patología producida por el SARS-CoV-2, virus de carácter zoonótico capaz de producir patologías multiorgánicas. La sintomatología que produce es variada. Ante la infección algunos pacientes presentan condiciones de gravedad. Los estudios han demostrado que el rol genético tiene gran afluencia sobre la respuesta ante la infección. El objetivo de investigación es detallar los genes que están implicados en la gravedad de la infección por SARS-CoV- 2. Metodología . Se realizó una revisión sistemática, con base a la búsqueda y análisis de artículos originales en bases de datos de alto impacto como PudMed, Google Académico, Scopus, Taylor & Francis, ProQuest SciencieDirect; se utilizaron operadores booleanos, criterios de inclusión y exclusión con el objetivo de obtener información precisa. Los genes de inmunidad como el HLA, ACE2 y TMPRSS2 están directamente involucrados con la gravedad de la infección por SARS-CoV- 2. Los polimorfismos de los genes del polyQ del receptor de andrógenos, factor ABO coadyuvan a un deterioro del estado patológico como consecuencia de la COVID-19. Conclusión. Los estudios demostraron que existen genes involucrados en la gravedad ante la infección de SARS -CoV-2, pues mencionan que los polimorfismos de los genes; HLA, del polyQ del receptor de andrógenos y factor ABO producen susceptibilidad ante el COVID-19. Así mismo los genes ACE2 y TMPRSS2 intervienen en el ingreso y expansión del virus. En las diversas razas existen variantes de los genes CCL2 y MBL lo que indica que algunas poblaciones son más susceptibles que otras.

Abstract COVID-19 is a pathology caused by SARS-CoV-2, a zoonotic virus capable of producing multi-organ pathologies. The symptomatology it produces is varied. Some patients present severe conditions after infection. Studies have shown that the genetic role has a great influence on the response to infection. Methodology . A systematic review was carried out, based on the search and analysis of original articles in high impact databases such as PudMed, Google Scholar, Scopus, Taylor & Francis, ProQuest SciencieDirect; Boolean operators, inclusion and exclusion criteria were used in order to obtain accurate information. Immunity genes such as HLA, ACE2 and TMPRSS2 are directly involved with the severity of SARS-CoV- 2 infection. Polymorphisms of androgen receptor polyQ genes, ABO factor contribute to a deterioration of the pathological state as a consequence of COVID-19. Conclusion . The studies demonstrated that there are genes involved in the severity of SARS-CoV-2 infection, since they mention that polymorphisms of the HLA, androgen receptor polyQ and ABO factor genes produce susceptibility to COVID-19. Likewise, ACE2 and TMPRSS2 genes intervene in the entry and expansion of the virus. In the different breeds there are variants of the CCL2 and MBL genes, which indicates that some populations are more susceptible than others.

Resumo COVID-19 é uma patologia causada pelo SARS-CoV-2, um vírus zoonótico capaz de produzir patologias multiorganismos. Os sintomas que ela produz são variados. Alguns pacientes se apresentam com condições severas após a infecção. Estudos demonstraram que o papel da genética desempenha um papel importante na resposta à infecção. O objetivo desta pesquisa é detalhar os genes que estão envolvidos na gravidade da infecção pelo SARS-CoV- 2. Metodologia. Foi realizada uma revisão sistemática, baseada na busca e análise de artigos originais em bancos de dados de alto impacto como PudMed, Google Scholar, Scopus, Taylor & Francis, ProQuest SciencieDirect; operadores booleanos, critérios de inclusão e exclusão foram utilizados para obter informações precisas. Resultados. Os genes de imunidade como HLA, ACE2 e TMPRSS2 estão diretamente envolvidos na gravidade da infecção pelo SARS-CoV- 2. Os polimorfismos dos genes receptores de androgênio polyQ, fator ABO contribuem para uma deterioração do estado patológico como conseqüência da COVID-19. Conclusão. Os estudos demonstraram que existem genes envolvidos na gravidade da infecção pelo SRA-CoV-2, pois mencionam que os polimorfismos dos genes HLA, androgênio receptor policarbonato e fator ABO produzem suscetibilidade à COVID-19. Os genes ACE2 e TMPRSS2 também estão envolvidos na entrada e propagação do vírus. Existem variantes dos genes CCL2 e MBL nas diferentes raças, indicando que algumas populações são mais suscetíveis do que outras.

Mechanism of B7-H3 molecule in renal clear cell carcinoma cell (786-O) metastasis / 中华微生物学和免疫学杂志

Objective:To investigate the effects of B7-H3 molecule on clear cell renal cell carcinoma (786-O) metastasis.Methods:Lentiviral transfection method was used to construct 786-O cells stably expressing low level of B7-H3 (shB7-H3 group) and a negative control cell line (shNC group). RT-qPCR, flow cytometry and Western blot were used to assess the efficiency of lentiviral transfection. CCK-8 method was used to detect the proliferation of 786-O cells in the two groups. Flow cytometry was performed to detect the changes in cell cycle. Cell scratch test and Transwell assay were used to detect the differences in cell migration and invasion. Western blot was used to detect the expression of marker proteins in the process of epithelial-mesenchymal transition (epithelial-mesenchymal transition, EMT). Changes in the expression of chemokines and their receptors were analyzed by flow cytometry and RT-qPCR. Effects of anti-CCL4 antibody on cell migration and invasion were analyzed by Transwell assay.Results:Flow cytometry showed that 786-O cells highly expressed B7-H3 molecules and the lentiviral transfection method successfully constructed the cell line with lower expression of B7-H3 (786-O-shB7-H3) and control cell line (786-O-shNC). B7-H3 molecule had no significant effect on the proliferation of 786-O cells. No significant difference in cell cycle was found between the two groups. Compared with 786-O-shNC cells, the migration and invasion ability of 786-O-shB7-H3 cells was suppressed. Moreover, the expression of EMT-related marker proteins (fibronectin and N-cadherin) was reduced and the expression of E-cadherin was increased in 786-O-shB7-H3 cells. The expression of CCL4 and its receptor CCR5 in the shB7-H3 group was lower than that in the shNC group. After intervention with anti-CCL4 antibody, the migration and invasion ability of 786-O-shNC cells was reduced, while that of 786-O-shB7-H3 cells had no significant change.Conclusions:Knocking down the expression of B7-H3 molecule had no significant effect on the proliferation of 786-O cells, but could affect the EMT process of 786-O cells and reduce tumor migration and invasion ability, thereby inhibiting tumor progression.

linc00941 regulates the proliferation, invasion and glycolysis of esophageal squamous cell carcinoma via miR-203/CCL2 axis / 中国肿瘤生物治疗杂志

@#[摘要] 目的：探究linc00941 作为ceRNA吸附miR-203 上调CC-趋化因子配体2（CC chemokine ligand 2，CCL2）的表达在食 管鳞癌（esophageal squamous cell carcinoma，ESCC）中的作用机制。方法：选取南阳医学高等专科学校第一附属医院58 例ESCC 患者的癌组织和癌旁组织，其中，男性患者33 例，年龄（49.3±18.6）岁，女性患者25 例，年龄（44.6±20.7）岁。qPCR 法检测 linc00941、miR-203、CCL2 在ESCC 组织和4 株人ESCC 细胞系（EC9706、KYSE30、ECA109 和TE1）以及人正常食管上皮细胞株 HET-1A细胞系中的表达。构建linc00941-wt、linc00941-mut、CCL2-wt、CCL2-mut 质粒并分别与miR-203 NC 或miR-203 模拟物 共转染到293T 细胞中。双荧光素酶报告基因实验验证linc00941、miR-203、CCL2 之间的相互作用。CCK-8 和Transwell 实验检 测细胞的增殖与侵袭能力。乳酸含量检测评价细胞的糖酵解能力。流式细胞术检测细胞的凋亡情况。糖酵解抑制剂2-DG以及 linc00941 共同干预ESCC细胞，以进一步观察linc00941 对ESCC细胞的调控作用。结果：在ESCC组织中和细胞系中linc00941、 CCL2 表达均上调，miR-203 表达下调（均P<0.05）。linc00941 与miR-203、miR-203 与CCL2 的相互作用在ECA109 细胞中得到证 实。下调linc00941 能够抑制ECA109 细胞的增殖、侵袭和糖酵解，并诱导细胞凋亡，该作用被miR-203 抑制剂部分逆转（均 P<0.05）。过表达CCL2 可以部分逆转敲减linc00941 对ECA109 细胞增殖、侵袭、糖酵解和凋亡的影响（均P<0.05）。结论： linc00941 能够吸附miR-203 进而上调CCL2 的表达，促进ESCC细胞的增殖、侵袭和糖酵解，诱导细胞凋亡。linc00941 对ESCC细 胞增殖、侵袭和凋亡的影响可能是通过调控糖酵解实现的。

Protective effect of Portulaca oleracea L. extract on acute liver injury induced by carbon tetrachloride in mice and its chemical composition analysis / 药学实践杂志

Objective To investigate the protective effect of the ethanol extract of Portulaca oleracea L. on acute liver injury induced by carbon tetrachloride in mice, and to analyze its effective components. Methods 80% ethanol purslane extract was centrifuged, vacuum distillated and vacuum dried into whole plant extract, supernatant extract and precipitated extract. Eighty ICR male mice were randomly divided into 8 groups: control group, liver injury model group, whole plant extract low-dose group, high-dose group, supernatant extract low-dose group, high-dose group, precipitation extract low-dose group, and high-dose group. After oral administration of distilled water or three kinds of purslane extract suspensions at different doses for 1 week, olive oil or CCl4 olive oil solution were injected subcutaneously respectively. After 16 hours, serum was collected to detect the levels of ALT, AST and IL-6 to evaluate the protective effect of purslane on acute liver injury. Ultra-high performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-Q-TOF/MS) was used to analyze the effective components of purslane extract. Results Compared with the model group, the levels of serum AST, ALT and IL-6 in high-dose whole plant extract group were significantly reduced. The serum ALT level of mice in the high-dose precipitation extract group was significantly reduced (P<0.05). The serum IL-6 level was decreased, but there was no significant difference. There were no significant changes in the levels of serum AST, ALT and IL-6 in the other intervention groups. 15 main components such as malic acid, citric acid, leucine, isoleucine, adenosine, succinic acid, genistein, tyrosine and phenylalanine were identified by UPLC-Q-TOF/MS. Conclusion Purslane whole plant ethanol extract has hepatoprotective and anti-inflammatory effects on CCl4 acute liver injury mice, which may be a combined effect of 15 active components.