[Process in the role of vasoactive intestinal peptide in the treatment of ulcerative colitis by regulating the balance of T follicular helper cells/T follicular regulatory cells].

Ulcerative colitis (UC) is an autoimmune disease based on the persistent damage of colonic mucosal barrier. It has been found that the abnormal expression of follicular helper T (Tfh) cells and follicular regulatory T (Tfr) cells is closely related to the occurrence and development of UC. Tfh cells can secrete pro-inflammatory factors and assist B cells to produce antibodies, which can promote the development of UC, while Tfr cells can inhibit the activity of Tfh cells and secrete anti-inflammatory factors. How to regulate the balance between them has become one of the potential therapeutic targets of UC. Vasoactive intestinal peptide (VIP) has preventive and therapeutic effect on UC, and its mechanism is closely related to the regulation of Tfh/Tfr cell balance, which can provide help for the treatment of UC.

Porous LaFeO3 perovskite catalysts synthesized by different methods and their high activities for CO oxidation.

In this study, spherical α-Fe2O3 prepared by the hydrothermal method was used as a template for the first time; LaFeO3 perovskite catalysts were successfully synthesized by the molten salt method (M-LF-T), sol-gel method (S-LF-T), and co-precipitation method (C-LF-T), respectively. To determine the optimal synthesis method, X-ray diffraction patterns were obtained and showed that single phase LaFeO3 with good crystallinity was prepared by the molten salt method after calcination at 600 °C for 4 h. SEM and TEM images showed that the M-LF-600 catalyst preserved the spherical structure of α-Fe2O3 template. Compared with the catalysts synthesized by the sol-gel method and co-precipitation method, the M-LF-600 catalyst had the highest BET surface area of 16.73 m2 g-1. X-ray photoelectron spectroscopy analysis showed that the M-LF-600 catalyst had the highest surface Fe3+/Fe2+ molar ratio and the best surface oxygen adsorption capacity. The CO oxidation of the LaFeO3 catalyst demonstrated that the M-LF-600 catalyst had the best catalytic performance.

Purification of recombinant human chemokine CCL2 in E. coli and its function in ovarian cancer.

Chemokine (CC-motif) ligand 2 (CCL2) is an inflammatory cytokine that regulates the infiltration and migration of monocytes. It is highly expressed by both tumor and stromal cells and has been associated with tumorigenesis. However, the effect of the exogenous administration of CCL2 on ovarian cancer remains largely unknown. In this report, we attempted to establish an expression system in Escherichia coli to produce recombinant hCCL2. The recombinant plasmid containing the hCCL2 cDNA was prepared using the prokaryotic-expression plasmid pGEX-5X-3 and transformed into E. coli BL21. GST-hCCL2 was successfully induced by 0.1 mmol/L IPTG at 20 °C for 6 h, and the recombinant protein was purified using affinity chromatography. The purified protein was identified by SDS-PAGE and Western Blot. In vitro experiments revealed that rhCCL2 promoted the proliferation of ovarian cancer cells and increased the levels of phosphorylation of MEK and ERK1/2, and the levels of JUN, RELB and NF-κB2 mRNA. Furthermore, inhibition of ERK signaling by treatment with PD98059 decreased ovarian cancer cell proliferation and levels of JUN, RELB, and NF-κB2 mRNA, indicating that exogenous rhCCL2 increased the proliferation of ovarian cancer cells, partially by activating the MAPK/ERK pathway, and by targeting JUN, RELB, and NF-κB2. Our study uncovered a promoting role of exogenous CCL2 on ovarian cancer cell proliferation through the MAPK/ERK signaling pathway, which may facilitate the discovery of more potential roles of CCL2 in ovarian cancer. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-020-02571-0.

Shen-Qi-Jie-Yu-Fang exerts effects on a rat model of postpartum depression by regulating inflammatory cytokines and CD4(+)CD25(+) regulatory T cells.

BACKGROUND: Shen-Qi-Jie-Yu-Fang (SJF) is composed of eight Chinese medicinal herbs. It is widely used in traditional Chinese medicine for treating postpartum depression (PPD). Previous studies have shown that SJF treats PPD through the neuroendocrine mechanism. AIM: To further investigate the effect of SJF on the immune system, including the inflammatory response system and CD4(+)CD25(+) regulatory T (Treg) cells. MATERIALS AND METHODS: Sprague Dawley rats were used to create an animal model of PPD by inducing hormone-simulated pregnancy followed by hormone withdrawal. After hormone withdrawal, the PPD rats were treated with SJF or fluoxetine for 1, 2, and 4 weeks. Levels of Treg cells in peripheral blood were measured by flow cytometry analysis. Serum interleukin (IL)-1ß and IL-6 were evaluated by enzyme-linked immunosorbent assay, and gene and protein expressions of IL-1RI, IL-6Rα, and gp130 in the hippocampus were observed by reverse-transcription polymerase chain reaction and Western blot. RESULTS: Serum IL-1ß in PPD rats increased at 2 weeks and declined from then on, while serum IL-6 increased at 1, 2, and 4 weeks. Both IL-1ß and IL-6 were downregulated by SJF and fluoxetine. Changes in gene and protein expressions of IL-1RI and gp130 in PPD rats were consistent with changes in serum IL-1ß, and were able to be regulated by SJF and fluoxetine. The levels of Treg cells were negatively correlated with serum IL-1ß and IL-6, and were decreased in PPD rats. The levels of Treg cells were increased by SJF and fluoxetine. CONCLUSION: Dysfunction of proinflammatory cytokines and Tregs in different stages of PPD was attenuated by SJF and fluoxetine through the modulation of serum concentrations of IL-1ß and IL-6, expressions of IL-1RI, and gp130 in the hippocampus, and CD4(+)CD25(+) Treg cells in peripheral blood.

[Effect of Yinao Jieyu Recipe on Brain Derived Neurotrophic Factor of the Limbic System in Post-Stroke Model Rats].

OBJECTIVE: To observe the evolutionary tendency of brain derived neurotrophic factor (BDNF) of the limbic system in post-stroke model rats and the intervention effect of Yinao Jieyu Recipe (YJR). METHODS: Male Wistar rats were randomly divided into the normal control group (n =6), the sham-operation group (n =7), the multiple cerebral infarction (MCI) group (n =10), the post-stroke depression (PSD) group (n =10), the Chinese medicine (CM) treatment group (n =10), and the Western medicine (WM) treatment group (n =10) according to random digit table after open-field testing. Rats in the normal control group were routinely fed. 0. 3 mL normal saline was intravenously pushing from the external carotid artery to rats in the sham-operation group, and distilled water administered to them by gastrogavage. Each dose allogenic microthrombi were in vitro pushed to rats in the rest groups from the external carotid artery. The PSD model was duplicated by 21-day chronic unpredictable mild stress (CUMS) and single cage feeding in the PSD group 7 days after surgery. After preparing models rats in the CM group and the WM group were administered with YJR and Nimodipine respectively for 4 successive weeks. Changes of BDNF and the intervention effect of YJR were observed at week 1, 2, and 4 after intervention. RESULTS: Immunohistochemical results of BDNF showed, compared with the normal control group, expression levels of BDNF in the hippocampus, hypothalamus, and amygdala decreased in the MCI group at week 2 and 4 (P <0. 01 , P <0. 05) ; expression levels of BDNF in each part decreased in the PSD group at week 1-4 (P <0.01). Compared with the MCI group, expression levels of BDNF in each part decreased in the PSD group at week 1-4 (P <0. 01). Compared with the PSD group, expression levels of BDNF in each part increased in the CM group at week 1-4 (P <0. 01). CONCLUSION: BDNF changes existed in post-stroke model rats, and YJR could slow down this progress.

[Angiogenesis of brain after ischemia/reperfusion injury of brain in aged rats and changes in expressions of basic fibroblast growth factor and transformation growth factor-ß1].

OBJECTIVE: To elucidate the mechanism of angiogenesis after cerebral ischemia/reperfusion (I/R) in the aged rats by observing the changes in expressions of basic fibroblast growth factor (bFGF) and transformation growth factor-ß1 (TGF-ß1). METHODS: Young Sprague-Dawley (SD) rats were classified into groups by random digits table, and aged SD rats were stratified by different body weight. Rats were randomly divided into groups of sham operation, ischemia (I) 3 hours, I/R 1, 3, 6, 12 days, with 6 rats in each group. Focal cerebral I/R model was reproduced by intraluminal filament technique. Microvessel density (MVD) of brain tissue, sum area of lumens were observed, and the expressions of bFGF protein, TGF-ß1 protein and TGF-ß1 mRNA were assessed with immunohistochemistry and hybridization in situ. RESULTS: MVD in young model group began to increase at I 3 hours, peaking at I/R 6 days, maintained up to I/R 12 days. MVD in aged model group began to descend at I 3 hours and continued to I/R 12 days. Sum area of lumens in young model group increased markedly at I/R 1 day, gradually lowered at I/R 1-6 days, and increased obviously again at I/R 12 days. Sum area of lumens in aged model group reached peak at I/R 1 day, gradually decreased subsequently. MVD in aged sham operation group were higher than that in young sham operation group (6.88±1.60 vs. 5.50±1.53, P<0.01). MVD and sum area of lumens in aged model group at I/R 1, 3, 6, 12 days were lower than young model group. Expressions of bFGF protein, TGF-ß1 protein in young and aged model group were both gradually up-regulated, all of them reaching peak at I/R 3 days, and lowered gradually at I/R 3-12 days subsequently. Expressions of bFGF protein (grey level) in both aged sham operation group and those of model group at I 3 hours, I/R 1, 3, 12 days were lower than those of young sham operation and those of the model group at the same time points (176.80±5.10 vs. 172.82±1.53, 171.81±2.43 vs. 167.85±2.41, 167.99±5.51 vs. 164.90±2.15, 152.98±4.11 vs. 150.75±1.11, 165.67±3.55 vs. 161.73±1.29, P<0.05 or P<0.01). Expressions of TGF-ß1 protein (grey level) in both aged sham operation and those of model group at I 3 hours, I/R 1, 3, 6, 12 days were all lower than those of young sham operation and those of model group at the same time points (182.69±3.12 vs. 176.13±4.08, 176.89±2.30 vs. 170.56±7.47, 171.74±2.70 vs. 165.43±2.91, 157.17±5.20 vs. 150.43±4.28, 161.72±4.81 vs. 155.37±2.92, 167.69±2.18 vs. 160.28±3.59, all P<0.01). TGF-ß1 mRNA expressions in both young model group and aged model group reached peak at I/R 1 day, gradually lowered subsequently. Expressions of TGF-ß1 mRNA (gray level) in both aged sham operation and those of model group at I 3 hours, I/R 3, 6, 12 days were lower than those of young sham operation and also model group at the same time points (176.51±9.52 vs. 169.09±5.08, 176.75±5.74 vs. 165.36±4.78, 177.33±5.68 vs. 165.25±8.14, 178.46±4.91 vs. 170.51±4.29, 203.95±4.51 vs. 181.98±5.59, all P<0.01). CONCLUSION: Angiogenesis is obviously weak after cerebral I/R in the aged, and the mechanism of which might be related to the down-regulation of expressions of bFGF protein, TGF-ß1 protein and TGF-ß1 mRNA. Aging factor may be one of the main reasons which induce the down regulation of expressions mentioned above.

Overexpression of p27(KIP1) induced cell cycle arrest in G(1) phase and subsequent apoptosis in HCC-9204 cell line.

AIM:We have previously reported that inducible over-expression of Bak may prolong cell cycle in G(1) phase and lead to apoptosis in HCC-9204 cells. This study is to investigate whether p27(KIP1) plays an important role in this process.METHODS:In order to elucidate the exact function of p27(KIP1) in this process, a zinc inducible p27(KIP1) stable transfectant and transient p27(KIP1)-GFP fusion transfectant were constructed. The effects of inducible p27(KIP1) on cell growth, cell cycle arrest and apoptosis were examined in the mock, control pMD vector, and pMD-KIP1 transfected HCC-9204 cells.RESULTS:This p27(KIP1)-GFP transfectant may transiently express the fusion gene. The cell growth was reduced by 35% at 48 h of p27(KIP1) induction with zinc treatment as determined by trypan blue exclusion assay. These differences remained the same after 72h of p27(KIP1) expression. p27(KIP1) caused cell cycle arrest after 24 h of induction, with 40% increase in G(1) population. Prolonged p27(KIP1) expression in this cell line induced apoptotic cell death reflected by TUNEL assay. Fourty-eight h and 72 h of p27(KIP1) expression showed a characteristic DNA ladder on agarose gel electrophoresis.CONCLUSION:Bak may induce cell cycle arrest in G(1) phase through upregulating expression of p27(KIP1) and subsequently lead to apoptosis in HCC-9204 cells. The p27(KIP1) -GFP fusion protein can be transiently expre-ssed in HCC-9204 cells. The inducible p27(KIP1)-expressing cell line provides a model to assess p27(KIP1) function.