Anti-cancer Effect of Traditional Chinese Medicine in Regulating Cellular Senescence： A Review / 中国实验方剂学杂志

In recent years， the incidence and mortality rates of cancer have been increasing， posing a serious threat to human health. Western medicine mainly uses treatments such as surgical resection， chemotherapy， immunotherapy and targeted therapy， but they are prone to complications， drug resistance and adverse reactions. A growing number of studies have shown that traditional Chinese medicine has obvious advantages in the treatment of cancer， reducing the recurrence rate of cancer and improving the quality of survival of patients. Cellular senescence refers to a state of irreversible cell cycle growth arrest when cells cease to proliferate after a limited number of divisions， resulting in a decline in cell proliferation and differentiation capacities and physiological functions， accompanied by morphological changes such as flattening and multinuclear morphology. At the molecular level， it shows increased expression of DNA damage-related genes， reduced expression of cell cycle-related factors and significant secretory activity. The malignant development of cancer is closely related to cellular senescence. With the increasing number of cancer cell proliferation， cancer-related genes undergo continuous mutations， freeing them from cellular senescence and thus achieving unlimited proliferation. Through recent studies， it has been found that induction of tumor cell senescence， possibly through modulation of cellular DNA damage， cell cycle arrest and senescence-associated secretory phenotype （SASP）， which converts the suppressive immune tumor microenvironment to an activated immune tumor microenvironment and thus reverses the escape of tumor cell senescence， is a promising strategy for cancer therapy. However， the mechanism of cellular senescence in cancer progression is not fully understood， especially the anti-cancer role played by traditional Chinese medicine in regulating cellular senescence. This article summarized and concluded the specific molecular mechanisms of cellular senescence， the role of cellular senescence in cancer progression， and the mechanism of anti-cancer effects of traditional Chinese medicine based on cellular senescence from the perspective of regulating cellular senescence， with a view to providing ideas and methods for the anti-cancer effects of traditional Chinese medicine and the development of new drugs.

Improvement effects of Runchang granules on the constipation in mice and its mechanism / 中国药房

OBJECTIVE To investigate the improvement effects of Runchang granules on the constipation in mice and its potential mechanism. METHODS The mice were randomly divided into normal control group， model group， Runchang granules low-dose and high-dose groups （5， 10 g/kg）， mosapride group （0.003 g/kg， positive control）， with 6 mice in each group. The latter 4 groups were given loperamide intragastrically （0.004 g/kg）， twice a day， for 3 consecutive days. Normal control group and model group were given purified water intragastrically， and administration groups were given relevant medicine intragastrically for 7 consecutive days. After the last medication， fecal moisture content and intestinal motility of mice were determined， while the structures of colon and ileum， and the secretion of colonic mucus were observed. Protein expressions of tyrosine kinase receptor （c-kit）， mucin 2 （MUC2） and stem cell factor （SCF） were determined in colon； meanwhile， the mRNA expression levels of inflammatory factors [tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， IL-1β， inducible nitric oxide synthase （iNOS）] as well as factors related to promoting intestinal motility [neuronal nitric oxide synthase （nNOS）， smooth muscle myosin light chain kinase （smMLCK）， 5-hydroxytryptamine 4 receptor （5-HT4R）， MUC2， SCF， c-kit] were determined. RESULTS Compared with model group， the fecal water content， intestinal propulsion rate， protein expression of c-kit in colon， relative expressions of MUC2 and SCF protein， and mRNA expressions of factors related to promoting intestinal motility （except for nNOS and SCF in Runchang granules low-dose group） were all increased significantly in Runchang granules low-dose and high-dose groups， and mosapride group （P＜0.05 or P＜0.01）. mRNA expression levels of inflammatory factors were decreased significantly（P＜0.05 or P＜0.01）. Both colon and ileum injuries improved， and the secretion of colon mucus was increased significantly in Runchang granules high-dose group （P＜0.01）. CONCLUSIONS Runchang granules have laxative effect and can improve constipation in mice， and its mechanism may be related to the promotion of the secretion of colon mucus and MUC2 expression， and the activation of SCF/c-kit signaling pathway.

Effect of moxibustion on immune function homeostasis in rats with diarrhea irritable bowel syndrome based on SCF/c-kit signaling pathway / 中国针灸

OBJECTIVE@#To observe the effects of moxibustion on the stem cell factor (SCF)/tyrosine kinase receptor (c-kit) signaling pathway and immune function in rats with diarrhea irritable bowel syndrome (IBS-D), and to explore the mechanism of moxibustion for IBS-D.@*METHODS@#Among 52 young rats born from 6 healthy pregnant SPF rats, 12 rats were randomly selected into the normal group, and the remaining 40 rats were treated with the three-factor combination method of maternal separation, acetic acid enema and chronic restraint stress to establish the IBS-D rat model. Thirty-six rats with successful IBS-D model were randomly divided into a model group, a moxibustion group, and a medication group, 12 rats in each group. The rats in the moxibustion group were treated with suspension moxibustion at "Tianshu" (ST 25) and "Shangjuxu" (ST 37); the rats in the medication group were treated with intragastric administration of rifaximin suspension (150 mg/kg). All the treatments were given once a day for 7 consecutive days. The body mass, loose stool rate (LSR), the minimum volume threshold when abdominal withdrawal reflex (AWR) scored 3 were measured before acetic acid enema (35 days old), after modeling (45 days old), and after intervention (53 days old). After intervention (53 days old), HE staining was used to observe the morphology of colon tissue, and spleen and thymus coefficients were measured; ELISA method was used to detect serum inflammatory factors (tumor necrosis factor a [TNF-a], interleukin [IL]-10, IL-8), T-lymphocyte subsets (CD+4, CD+8, CD+45), value of CD+4/CD+8 and immune globulin (IgA, IgG, IgM); real-time PCR method and Western blot method was used to detect the expression of SCF, c-kit mRNA and protein in colon tissue; immunofluorescence staining method were used to detect positive expression of SCF and c-kit.@*RESULTS@#After intervention, compared with the normal group, in the model group, the body mass and the minimum volume threshold when AWR scored 3 were decreased (P<0.01), LSR, spleen and thymus coefficients, serum levels of TNF-α, IL-8, CD+4, CD+45, CD+4/CD+8, IgA, IgG, IgM were increased (P<0.01), serum IL-10 level and protein and mRNA expression of SCF and c-kit in colon tissue were decreased (P<0.01), and the positive expression of SCF and c-kit was decreased (P<0.01). Compared with the model group, in the moxibustion group and the medication group, the body mass and the minimum volume threshold when AWR scored 3 were increased (P<0.01, P<0.05), LSR, spleen and thymus coefficients, serum levels of TNF-α, IL-8, CD+4, CD+8, CD+45, CD+4/CD+8, IgA, IgG, IgM were decreased (P<0.01, P<0.05), serum IL-10 level and protein and mRNA expression of SCF and c-kit in colon tissue were increased (P<0.01), and the positive expression of SCF and c-kit was increased (P<0.01). Compared with the medication group, in the moxibustion group, the level of serum CD+4 was decreased (P<0.05), the value of CD+4/CD+8 was increased (P<0.01), and there was no significant difference in other indexes (P>0.05). The expression of SCF and c-kit mRNA was positively correlated with the minimum volume threshold when AWR scored 3 and IL-10 (P<0.01), and negatively correlated with remaining indexes (P<0.01, P<0.05).@*CONCLUSION@#Moxibustion could reduce visceral hypersensitivity, improve symptoms of abdominal pain and diarrhea in IBS-D rats, and its mechanism may be related to up-regulation of the expression of SCF/c-kit signaling pathway and improvement of IBS-D immune function.

Effect of stem cell factor on the angiogenic ability of cocultured DPSCs and HUVECs / 口腔疾病防治

Objective@#To study the effect of stem cell factor (SCF) on the angiogenic ability of cocultured dental pulp stem cells (DPSCs) and human umbilical vein endothelial cells (HUVECs).@*Methods @#This study has been reviewed and approved by the Ethics Committee. The experiment was split into the HUVECs, SCF+HUVECs, DPSCs+HUVECs, and SCF+DPSCs+HUVECs groups. A mixture of SCF and culture medium was used to prepare a mixed culture medium with an SCF concentration of 100 ng/mL. In vitro coculture of DPSCs and HUVECs was performed at a 1∶5 ratio. CCK-8 proliferation assay was used to observe the proliferative capacity of cells in each group on days 1, 3, 5, and 7. Wound healing and Transwell migration assays were used to detect the effect of SCF on cell migration under either direct or indirect coculture conditions, respectively. In vitro angiogenesis experiments were performed to detect the angiogenic capacity of the cells in each group. The vascular endothelial growth factor A (VEGFA) concentration in the cell culture supernatant was detected using ELISAs, and the protein expression levels of CD31, CD34, and VEGFA were detected using Western blot analysis. @*Results @# Wound healing and Transwell migration experiments showed that SCF significantly promoted the migration of cocultured DPSCs and HUVECs (P<0.05). The in vitro angiogenesis experiment showed that the number of branches and the total length of branches of tubular structures in the SCF+DPSCs+HUVECs group were significantly greater than those of the other groups (P<0.05), and the expression levels of the vascular-related proteins CD31, CD34, and VEGFA in this group were greater (P<0.01). @*Conclusion @# SCF can enhance the migration and in vitro angiogenesis of cocultured DPSCs and HUVECs.

Increased tumor-associated mast cells facilitate thyroid cancer progression by inhibiting CD8+ T cell function through galectin-9

As an important component of solid tumors, mast cells show specific phenotypes in various tumor microenvironments. However, the precise mechanism of mast cell accumulation and the phenotypic features of thyroid cancer (TC) remain largely unknown. Here, we found that mast cells were obviously recruited to tumor tissue by TC-derived stem cell factor (SCF). With tumor progression, mast cell levels increased gradually. In addition, intratumoral mast cells expressed higher levels of the immunosuppressive molecule galectin-9, which effectively suppresses CD8+ T-cell antitumor immunity in vitro. Blocking galectin-9 on tumor-infiltrating mast cells reversed the immunosuppression of CD8+ T cells. In conclusion, our data elucidated novel protumorigenic and immunosuppressive roles of mast cells in TC. In addition, our results indicated that blocking mast cells may impede tumor progression and ameliorate the prognosis of TC patients.

EHD2 Affects the Proliferation of Esophageal Squamous Cell Carcinoma by Regulating the Cyclin Dl-CDK4-pRb Signaling Axis / 中国生物化学与分子生物学报

Actin-binding proteins (ABPs) are important components of the F-actin cytoskeleton and affect the dynamics of F-actin by promoting the polymerization and depolymerization of actin. Numerous studies have shown that F-actin and actin-binding proteins are involved in all stages of carcinogenesis. Our analysis of esophageal carcinoma proteomic data showed that the actin-binding protein EHD2 (E p s l 5 homology domain-containing protein 2) is expressed at low levels in esophageal squamous cell carcinoma tissues and patients with lower EHD2 expression had poorer prognosis. Previous studies have revealed that EHD2 is involved in the regulation of glucose metabolism, autophagy and tumor cell migration. However, the role and mechanism of EHD2 in esophageal squamous cell carcinoma progression remain unclear. This study aimed to explore the effect of EHD2 on the proliferation of esophageal squamous cell carcinoma. Immunofluorescence and cell fractionation analysis showed that EHD2 was not only localized in the cell membrane and cytoplasm, but also in the nucleus. Colony formation, EdU labeling and flow cytometry were used to determine the effect of EHD2 on the proliferation of esophageal squamous cell carcinoma. The results showed that overexpression of EHD2 and EHD2-3×NLS (nuclear localization signal) inhibited proliferation, cell cycle G

MiR-654-5p Inhibited Invasion and Metastasis of Breast Cancer Cells by HCFC1 / 中国生物化学与分子生物学报

Studies have confirmed that microRNA (miRNAs) is involved in the development and progression of tumors by targeting multiple genes. However, the molecular mechanism of miR-654-5p in in- hibiting the invasion and metastasis of breast cancer cells through the targeted regulation of host cell factor 1 (HCFCl) is still unclear. The analysis of bioinformatics datasets found that miR-654-5p was downregu-lated in breast cancer tissues and was associated with poor prognosis (P = 0. 013). Quantitative real-time PCR (Quantitative real-time PCR, qRT-PCR) showed that the expression of miR-654-5p in MDA-MB-231 cells was decreased (P<0. 05), and the expression of miR-654-5p was significantly increased after transfection of the overexpressed plasmid (P<0. 05) as compared with the control group. The 5-Ethynyl-2'-deoxyuridine (EdU) proliferation experiment and Transwell assay showed that overexpression of miR-654-5p inhibited the proliferation, migration and invasion of MDA-MB-231 cells (P<0. 05). Hub gene HCFCl of miR-654-5p was screened and constructed by Cytoscape software, and it was found that miR-654-5p was negatively correlated with the expression of HCFCl. The expression of HCFCl was increased in breast cancer tissues and closely correlated with lymph node metastasis, and patients with high expression of HCFCl had poor prognosis (P = 0. 0039). Dual-luciferase assay confirmed that miR-654-5p could bind to the 3'-UTR of HCFClmKNA (P<0. 05). Western blot results showed that compared with human normal breast epithelial cells MCF-10A, the expression of HCFCl was increased in MDA-MB-231 cells (P<0. 05), and the expression of HCFCl was significantly down-regulated after overexpression of miR-654-5p (P<0. 05). Transwell experiment results showed that the migration and invasion ability of MDA-MB-231 cells after overexpression of miR-654-5p was significantly decreased compared with the control group. Co-transfection of HCFCl can partially reverse the inhibitory effect of miR-654-5p on the migration and invasion ability of breast cancer cells (P<0. 05). In conclusion, miR-654-5p can inhibit the proliferation, invasion and metastasis of breast cancer cells by regulating the expression of HCFCl.

Construction of immunodeficient mouse models with long-term expression of human cytokines based on piggyBac transposon system / 吉林大学学报(医学版)

Objetive: To investigate the long-term expression of the piggyback (PB) transposon system expressing human interleukin-6 (IL-6), nterleukin-3 (IL-3), interleukin-15 (IL-15), stem cell factor (SCF) and granulocyte-macrophage ctimulang Factor (GM-CSF) genes in the immunodeficient mice, and to provide a simple, long-term and new method for improving the reconstruction of human immune cells in the humanized mouse models. Methods: The PB transposon plasmid (PB-GFP) containing the GFP gene was constructed, and the PB transposon plasmid (PB-5F) contaning human IL-6, IL-3, IL-15, SCF, and GM-CSF genes was constructed. The 293T cells were divided into negative control group (non-transfection), postive control group (transfected with pLVTHM), transient transfection group transfected with PB-GFP plasmid and stable transfection group transfected with PB-GFP plasmid together with transposase plasmid (super-PB)]. The proportions of GFP cells in varions groups were measured by flow cytometry every three days after transfection. The NOD. Cg-Prkdcscid IL2rgun1wj1/SzJ (NCG) mice were divided into transient transfection group and stable transfection group. The mice in transient transfection group were transfected with PB-GFP alone, and the mice in stable transfection group were transfected with PB-5F and super-PB. On the 1st, 4th, 5th and 9th days and the followng every week after the transfection, the blood samples were collected, and the serum was separed; the levels of IL-6, IL-3, IL-15, SCF, and GM-CSF in serum of the mice in various groups were detected by ELISA Results: At 30 d after transfection of PB-GFP, the percentage of GFP1 cells of the mice in stable transfection group (4. 61% + 0.42%) was significantly higher than those in postive control group (0.58% +0.05%) and transient transfection group (0.86%+ 0.10%) (P<0.05). At 30 d after transfection of PB-5F plasmid, the levels of serum IL-6, IL-15 and GM-CSF of the NCG mice in stable transfection group were significantly higher than those in transient transfection group (P

Study on the significance of cytokine in bronchoalveolar lavage fluid in children with refractory mycoplasma pneumoniae pneumonia and the correlation with serum C reactive protein / 国际儿科学杂志

Objective To explore the levels and clinical significance of MCP-4,IL-25,TNF-α and CysLTR-1 in bronchoalveolar lavage fluid (BALF) of children with refractory mycoplasma pneumoniae pneumonia (RMPP) and their correlation with serum C-reactive protein (CRP).Methods One hundred and nine children diagnosed as RMPP who underwent fiberoptic bronchoscopy in acute stage (course of disease within 2 weeks) were selected as the experimental group.According to the manifestations of mucosa,secretions and lumen under bronchoscope,the patients were divided into RMPP1 group (68 cases of severe pathological injury under bronchoscope) and RMPP2 group (41 cases of mild pathological injury under bronchoscope).They were divided into RMPP1 wheezing group (20 cases),RMPP1 non-wheezing group (48 cases),RMPP2 wheezing group (15 cases) and RMPP2 non-wheezing group (26 cases).15 children with non-mycoplasma pneumoniae pneumonia (NMPP) and non-wheezing lobar pneumonia in the same period were selected as control group 1.At the same time,15 children without pneumonia underwent bronchial foreign body (FB) removal as control group 2.The levels of MCP-4,IL-25,TNF-α and CysLTR-1 in BALF of children in experimental group were determined by double antibody sandwich ELISA.Serum CRP,D dimer (DD),ALT and peripheral blood neutrophil percentage (N％) were also detected.Results (1) The levels of CRP,DD,ALT and N％ in RMPP1 group with severe bronchoscopic manifestations were higher than those in RMPP2 group with relatively mild bronchoscopic manifestations (all P ＜ 0.05).(2) The mean levels of IL-25 (117.8 ng/L),TNF-α (26.01ng/L),CysLTR-1 (0.71 ng/L) and MCP-4 (53.38 ng/L) in RMPP1 wheezing group were higher than those in the other five groups (P ＜ 0.05).The mean levels of IL-25 (85.79 ng/L),TNF-α (19.2 ng/L),CysLTR-1 (0.59 ng/L) and MCP-4 (44.16ng/L) cells in RMPP2 wheezing group were higher than those in RMPP2 non-wheezing group,NMPP group and FB group (all P ＜0.05).There was no statistical difference between the other two groups (P ＞ 0.05).(3) CRP was positively correlated with IL-25,MCP-4 and TNF-a (all P ＜ 0.05),but not with CysLTR-1.Conclusion (1) Clinical laboratory indicators such as CRP,DD,ALT and N％ can assist in early identification of RMPP.The higher the above indicators,the more serious performance of RMPP under microscope.(2) Cytokines MCP-4,IL-25,CysLTR-1 and TNF-α all participate in the pathogenesis of RMPP,and may play an important role in the occurrence of wheezing and development of asthma in children induced by MP infection.(3) Serum CRP levels were positively correlated with the levels of IL-25,MCP-4 and TNF-α in BALF of RMPP wheezing children.Both MCP-4 and IL-25 selectively affected Th2-induced Th2 cells.CRP is associated with IL-25 and MCP-4 who mediated immune inflammation injury.It is speculated that CRP may also cause Th2-mediated immune inflammation injury by affecting Th2 cells.

Correlation between microRNA and rheumatoid arthritis activity / 中华风湿病学杂志

Objective To investigate the correlation between miRNAs and clinical manifestations, laboratory examinations, serum cytokines and disease activity scores in rheumatoid arthritis (RA). Methods Thirty-eightnewly diagnosed RA patients who visited the department of Rheumatology of Inner Mongolia Medical Affiliated Hospital from October 2015 to December 2015 were enrolled in the study. Thirty-eight age and sex matched healthy volunteers were enrolled as the control group. MiR-125b, miR-21, miR-155, miR-346, miR-223, miR-146a were tested by real-time fluorescence quantitative polymerase chain reaction (real-time PCR). Datawere collected at the baseline and 3 weeks after the study. Solutions statistical package for the social sciences (SPSS) 20.0 version was usedfor statistical analysis. Data were analyzed using t test or χ2 test and Pearson test was used for correlation analysis.Levene method was used to test homogeneity of variance. Results MiR-125b (1.55±0.24), miR-155 (3.1±0.5), miR-346 (650±51), miR-223-3p (1.26±0.18), miR-146a-5p (2.39±0.25) levels in the RA group were significantly higher than those in the healthy control group at the baseline (0.84±0.16, 1.4±0.5, 304±101, 0.58±0.11, 1.09±0.27; t=15.36, 18.60, 18.77, 19.67, 21.66; P<0.05). MiR-21-5p (0.91±0.09) was lower than the control group (1.51±0.21; t=-16.029, P<0.05), while miR-146a-3p, miR-21-3p, miR-223-5p showed no difference between the two groups. MiR-155 and miR-146a was positively correlated with RA disease activity scores (P<0.05). MiR-21 was negatively correlated with disease activity scores and serum cytokines including IL-1β, IL-6, IL-17a, IL-23 and Tumor necrosis factor (TNF)-α. After 3 months treatment, tenderness joint count, swelling joint count, laboratory tests and imaging examination were improved (P<0.05), while miRNAslevels in RA were decreased significantly respectively (P<0.05), mir-21 was increased significantly. Conclusion MiR-125b, miR-21, miR-155, miR-346, miR-223, miR-146a in RA are characteristically expressed in RA and related with the disease activity and treatment response. MiRNAs may hold a great promise as the diagnostic and prognostic biomarkers in RA.

Study on the significance of cytokine in bronchoalveolar lavage fluid in children with refractory mycoplasma pneumoniae pneumonia and the correlation with serum C reactive protein / 国际儿科学杂志

Objective@#To explore the levels and clinical significance of MCP-4, IL-25, TNF-α and CysLTR-1 in bronchoalveolar lavage fluid(BALF)of children with refractory mycoplasma pneumoniae pneumonia(RMPP)and their correlation with serum C-reactive protein(CRP).@*Methods@#One hundred and nine children diagnosed as RMPP who underwent fiberoptic bronchoscopy in acute stage(course of disease within 2 weeks)were selected as the experimental group.According to the manifestations of mucosa, secretions and lumen under bronchoscope, the patients were divided into RMPP1 group(68 cases of severe pathological injury under bronchoscope)and RMPP2 group(41 cases of mild pathological injury under bronchoscope). They were divided into RMPP1 wheezing group(20 cases), RMPP1 non- wheezing group(48 cases), RMPP2 wheezing group(15 cases)and RMPP2 non-wheezing group(26 cases).15 children with non-mycoplasma pneumoniae pneumonia(NMPP)and non-wheezing lobar pneumonia in the same period were selected as control group 1.At the same time, 15 children without pneumonia underwent bronchial foreign body(FB)removal as control group 2.The levels of MCP-4, IL-25, TNF-α and CysLTR-1 in BALF of children in experimental group were determined by double antibody sandwich ELISA.Serum CRP, D dimer(DD), ALT and peripheral blood neutrophil percentage(N%)were also detected.@*Results@#(1)The levels of CRP, DD, ALT and N% in RMPP1 group with severe bronchoscopic manifestations were higher than those in RMPP2 group with relatively mild bronchoscopic manifestations(all P<0.05). (2)The mean levels of IL-25(117.8 ng/L), TNF-α(26.01ng/L), CysLTR-1(0.71 ng/L)and MCP-4(53.38 ng/L)in RMPP1 wheezing group were higher than those in the other five groups(P<0.05). The mean levels of IL-25(85.79 ng/L), TNF-α(19.2 ng/L), CysLTR-1(0.59 ng/L)and MCP-4(44.16ng/L)cells in RMPP2 wheezing group were higher than those in RMPP2 non-wheezing group, NMPP group and FB group(all P<0.05). There was no statistical difference between the other two groups(P>0.05). (3)CRP was positively correlated with IL-25, MCP-4 and TNF-a(all P<0.05), but not with CysLTR-1.@*Conclusion@#(1)Clinical laboratory indicators such as CRP, DD, ALT and N% can assist in early identification of RMPP.The higher the above indicators, the more serious performance of RMPP under microscope.(2)Cytokines MCP-4, IL-25, CysLTR-1 and TNF-α all participate in the pathogenesis of RMPP, and may play an important role in the occurrence of wheezing and development of asthma in children induced by MP infection.(3)Serum CRP levels were positively correlated with the levels of IL-25, MCP-4 and TNF-α in BALF of RMPP wheezing children.Both MCP-4 and IL-25 selectively affected Th2-induced Th2 cells.CRP is associated with IL-25 and MCP-4 who mediated immune inflammation injury.It is speculated that CRP may also cause Th2-mediated immune inflammation injury by affecting Th2 cells.

Expression and significance of stem cell factor in nephridial tissue of rat model with uric acid nephro-pathy / 中华风湿病学杂志

Objective To investigate the association between stem cell factor (SCF) expression and tubulointerstitial fibrosis in the kidney of rat model with uric acid nephropathy. Methods Thirty-six Wistar rats were randomly divided into model group and control group. The rats in the model group were fed with adenine by lavage at a dose of 150 mg·kg-1·d-1, and the rats in the control group were fed with normal saline by lavage at equal volume. Six rats from each group were sacrificed respectively at week 4, 8 and 12. The mRNA levels of SCF and ColⅠin the kidney were detected by real-time fluorescence quantitative polymerase chain reaction (PCR), and their protein levels and infiltrated Mast cell (MC) were measured by immunohistochemistry. The difference and correlation of each index among different time points and groups were compared. The differences between the two groups were tested by t-test, multiple data were tested by one-way analysis of variance (ANOVA) and the correlation was analyzed by Pearson's correlation. Results ①The serum uric acid (SUA) [week 4, 8, 12: (302 ±41))μmol/L, (424 ±61) μmol/L, (518 ±57) μmol/L], creatinine[week 4, 8, 12:(151±9)μmol/L, (219±15)μmol/L, (299±21)μmol/L], urea nitrogen [week 4, 8, 12:(26.7±3.7) mmol/L, (40.3 ±5.7) mmol/L, (61.9 ±9.4) mmol/L], urine protein/creatinine (Up/Ucr) [week 4, 8, 12: (0.71 ±0.10) mmol/L, (1.18 ±0.11) mmol/L, (1.78 ±0.13) mmol/L] and the expressions of SCF mRNA [week 4, 8, 12: (1.19 ± 0.41), (1.69 ±0.63), (2.21 ±0.97)} and SCF protein [week 4, 8, 12: (1.42 ±0.33), (6.02 ±1.81), (10.03 ±2.69)] in nephridial tissue of model group's rats were significantly higher compared to the control group (all of t value>4.59, P<0.01), and the indexes were increasing gradually as the lavage going on. ②The expression of SCF was correlated with collagenⅠ, degree of renal interstitial injury, urine nitrogen, serum creatinine and UP/Ucr(At week 4, r=0.53, 0.42, 0.40 and 0.51 respectively;at week 8, r=0.60, 0.59, 0.41 and 0.39 respectively;at week 12, r=0.74, 0.61, 0.56 and 0.39 respectively, all of P value<0.05). ③ Compared with control group, the number of infiltrated MC was significantly higher in the model group, and was positively correlated with the expression of SCF (r=0.91, P<0.01). Conclusion Compared with the control group, the expression of SCF and the number of infiltrated MC in the renal interstitium are evidently increased, and are increasing gradually as the lavage going on and the deteriorating of renal interstitial damage. These results suggest that both may play important roles in the occurrence and development of uric acid nephropathy.

Phenotypic changes of interstitial cells of Cajal after intestinal obstruction in rat model

The objective was to study the effect of mechanical intestinal obstruction in rats on the phenotype of interstitial cells of Cajal (ICC). Healthy Wistar rats were randomly divided into sham-operation group (C), one day obstruction group (M1), two days obstruction group (M2), and three days obstruction group (M3), with 10 rats in each group. The expression of SCF mRNA and c-Kit protein in intestinal tissue was investigated by RT-PCR and immunohistochemistry. Compared with the sham-operation group, the relative expression of SCF mRNA and the expression of c-Kit protein in intestinal tissue were significantly decreased in both obstruction groups. Levels decreased gradually with the prolongation of obstruction time, and significantly decreased on the 3rd day after obstruction (P<0.05). Immunohistochemical staining of the small intestine showed that the number of ICC in the sham-operation group was the highest, and they were gradually decreased with the extension of obstruction time in the M1 to M3 groups. There was a significant difference between groups (P<0.05). Intestinal obstruction caused a decrease in the concentrations of SCF mRNA and c-Kit protein in ICC. With the prolongation of intestinal obstruction, the number of ICCs gradually decreased.

No detection of c-kit gene mutations in exons 9, 11, 13 and 17 and low CD117 expression in plaque-stage mycosis fungoides

Abstract: The growth factor receptor c-kit (CD117) is expressed in immature T-cells and in some advanced forms of mycosis fungoides. c-kit gene mutation results in unrestricted neoplastic proliferation. We aimed to detect by PCR the most frequent exon mutations in seventeen plaque-stage MF patients, in their perilesional skin and in healthy skin donors. We secondarily evaluated CD117 expression by immunohistochemistry in plaque-stage and tumor-stage MF. We detected no mutation in c-kit gene and low CD117 expression was confirmed on atypical cells in one patient. Complete c-kit exon and intron sequences should be assessed and more sensitive sequencing method could be also applied.

Progress in the study of stem cell factor receptor c-kit in skin melanin metabolism / 中华整形外科杂志

Skin melanin metabolism is a complex and fine regulation process, which is controlled by multiple genes, many enzymes and proteins. With the in-depth study of the factors affecting the skin melanin metabolism, scientists have found that the signaling pathway initiated by the combination of stem cell factor receptor c-kit and its ligand stem cell factor plays an important role in the metabolism of skin melanin.The paper summarizes the recent research progress on the role of c-kit receptor in the skin melanin metabolism, including the molecular mechanism and the clinical application.

Cultivation and identification of mouse peritoneal and bone marrow-derived mast cells / 中华皮肤科杂志

Objective To explore the in vitro culture methods for oriented differentiation of peritoneal cells and bone marrow cells into high-purity mast cells,and to identify the function of these mast cells.Methods Peritoneal cells and bone marrow cells were isolated from the peritoneal cavity lavages and femur of C57BL/6 mice,and cultured with both interleukin-3 (IL-3) and stem cell factor for 2 and 4 weeks respectively.Light microscopy was performed to observe the morphology of these cells,toluidine blue staining to identify the degree of maturity of these mast cells,and flow cytometry to measure the expression of cell surface markers C D 117 and FceR Ⅰ α.After the stimulation with compound 48/80 at different concentrations,the degranulation rate of mast cells was counted under the microscope,and β-hexosaminidase release rate was measured by spectrophotometry.Results After 2-or 4-week culture,the mouse peritoneal and bone marrow cells all manifested as refractive suspension cells of uniform size.Toluidine blue staining showed violaceous metachromatic granules in the cytoplasm of the two kinds of cells.The proportions of CD117 or FcεR Ⅰ α single-positive peritoneal and bone marrow-derived mast cells were all more than 95％,and the proportions of CD117/FcεR Ⅰ α double-positive peritoneal and bone marrow-derived mast cells were 97.68％ ± 0.80％ and 96.12％ ± 0.76％ respectively.The degranulation rates of mast cells in the 100-and 1 000-mg/L compound 48/80 groups significantly differed from those in the blank control group (all P ＜ 0.01).Compared with the blank control group,the β-hexosaminidase release rates significantly increased in bone marrow-derived mast cells in the 100-mg/L compound 48/80 group and peritoneal mast cells in the 10-and 100-mg/L compound 48/80 groups (P ＜ 0.01 or 0.05).Conclusion IL-3 and stem cell factor can co-induce the directed differentiation and proliferation of mouse bone marrow stem cells and peritoneal cells,so as to harvest highnuritv mature degranulated mast cells,and lay a foundation for subsequent cell biology research.

Abnormal SCF-C-kit system in IBS-D rat and the correlation between SCF-C-kit system and immune function / 中国药理学通报

Aim To explore the change of stem cell factor-C-kit (SCF-C-kit) system in irritable bowel syn-drome with diarrhea ( IBS-D) and the correlation be-tween SCF-C-kit system and immune dysfunction . Methods Twelve neonatal rats were divided into nor-mal group and model group with six rats in each group . IBS-D rat model was established through three-factor method ( mother-son separation , acetic acid stimulation and constraint ) . Immunohistochemistry was used to observe in situ protein expressions of SCF and C-kit. qPCR was used for mRNA expressions of SCF and C-kit.Correlation between SCF-C-kit system and spleen coefficients , thymus coefficients , TNF-α, IL-8 , IL-10 was analyzed by statistics .Results Compared with normal group , positive rates of SCF and C-kit protein in model group both decreased , and so did mRNA ex-pression .Expression of SCF was negatively correlative with spleen coefficient , TNF-αexpression and IL-8 ex-pression , while positively correlative with IL-10 expres-sion.Expression of C-kit was negatively correlated with thymus coefficient , spleen coefficient and TNF-α. Conclusion SCF-C-kit system of IBS-D is abnormal, which may be related with immune dysfunction .

The level and clinical significance of stem cell factor in lupus nephritis / 中国医师进修杂志

Objective To explore the relationship between the level of serum stem cell factor (SCF) and the activity of lupus nephritis (LN). Methods Sixty LN patients who had underwent routine renal biopsy were selected from March 2014 to September 2016. According to the systemic lupus erythematosus disease activity index (SLEDAI), the LN patients were divided into two groups: active nephritis group (30 cases) and stable nephritis group(30 cases). Meanwhile 30 healthy controls were selected as normal control group. Spearman correlation analysis test was used for correlations between the level of serum SCF and the activity of LN. Results The serum level of SCF was significantly higher in active nephritis group [(357.29 ± 63.85) ng/L] than that in stable nephritis group [(310.03 ± 40.17) ng/L] , the serum level of SCF in stable nephritis group was significantly higher than that in normal control group [(154.06 ± 22.49) ng/L], and there were significant differences (P0.05). Conclusions The level of SCF may play an important role in the occurrence and development of LN. It could reflect clinical and pathology changes and emerg as a potential serum biomarkers of LN.

Effect of meropenem on children with severe infection and treatment effect on the level of PCT / 中国生化药物杂志

Objective To explore the effect of meropenem on the treatment of children with severe infection and the effect on the level of PCT.MethodsThe clinical data of patients with infectious diseases treated in our hospital from December 2013 to February 2016 were retrospectively analyzed.According to the anti infective drugs were divided into control group and observation group, the control group was given routine antibiotic treatment, the observation group was given meropenem treatment.The treatment effect of the two groups were observed, the difference of serum PCT, cytokine level and blood gas indexes before and after treatment in the two groups were compared, correlation analysis with PCT level and serum levels of inflammatory cytokines and blood indexes of severe infection.ResultsIn the control group, 50 cases were successful, the observation group was successful in the treatment of the patients, there were no significant differences in the success rate between the two groups.Two groups of patients before treatment PCT and inflammatory cytokines level no difference, after treatment, the observation group PCT, IL-18, IL-6 and hs-CRP levels were lower than the control group;Two groups of children before treatment blood gas index no difference, after treatment, the observation group SpO2 level higher than the control group, P ETCO2 level was lower than the control group.PCT levels in children with severe infection were positively correlated with IL-18, IL-6, hs-CRP and P ETCO2 levels and negatively correlated with SpO2 levels.ConclusionMeropenem has better therapeutic effect in the treatment of children with severe infection, can significantly reduce the serum PCT level, has the value of clinical application.

Therapeutical effect of Toudingyikezhu extract on rats with nonalcoholic fatty liver and its mechanism / 吉林大学学报(医学版)

Objective:To observe the effect of Toudingyikezhu extract on the morphology of liver tissue and endoplasmic reticulum stress(ERS) of the rats with nonalcoholic fatty liver disease(NAFLD),and to clarify the possible mechanism of Toudingyikezhu of intervention of NAFLD.Methods:The NAFLD rat models were established by intraperitoneal injection of carbon tetrachloride solution combined with high-fat diet.A total 60 SD rats were randomly divided into control group,model group,low,middle and high doses of Toudingyikezhu extract groups,phosphatidylcholine group(n=10).The doses of Toudingyikezhu extracts were 0.50,1.00 and 2.00 g·mL-1,and the dose of phosphatidylcholine was 0.20 g·kg-1,4 weeks after administration,the body weights and liver wet weights were detected,and the liver indexes were calculated.The texture,elasticity,color and morphology of liver tissue of the rats were observed by HE staning.The fatty degeneration,inflammation and necrosis scores of liver tissue of the rats were calculated.The levels of serum tumor necrosis factor-α(TNF-α),interleukin-1 (IL-1) and interleukin-6(IL-6) of the rats were detected by ELISA method.Immunohistochemical SP method was used to determine the expression levels of GRP78 protein,and PCR method was used to determine the expression levels of GRP78 mRNA.Results:Compared with control group,the morphology of liver tissue of the rats in model group were abnormal under light microscope;the liver index,fatty degeneration,inflammation and necrosis scores of liver tissue of the rats in model group were increased(P<0.05),and the expression levels of GRP78 protein and mRNA were decreased(P<0.01).Compared with model group,the liver tissue pathology of the rats had different degrees of improvement;the fatty degeneration,inflammation and necrosis scores of liver tissue and the serum TNF-α,IL-1,IL-6 levels of the rats in different doses of Toudingyikezhu extract groups were decreased(P<0.05 or P<0.01);the expression levels of GRP78 protein and mRNA were decreased (P <0.05 or P<0.01).Conclusion:Toudingyikezhu extract may inhibit or block the ERS,reduce the synthesis of lipids and accelerate its decomposition in order to resist the injury of liver cells of the NAFLD rats by down-regulating the GRP78 expression.