Objective To investigate the relationship between cyclin A2 (CCNA2) and the prognosis of colon cancer, and its possible mechanism from the perspective of immune infiltration. Methods We downloaded the transcriptome data of colon cancer patients from The Cancer Genome Atlas database. Clinicopathological feature analysis and survival analysis were performed based on the expression levels of CCNA2. A total of 75 specimens of colon cancer and normal tissues were collected, and the expression level of CCNA2 was analyzed using immunohistochemical methods. Multivariate analysis was conducted to explore its relationship with clinicopathological features. Gene Set Enrichment Analysis (GSEA) was used to assess the potential molecular functions of CCNA2 in colon cancer. CIBERSORT algorithm was applied to calculate the correlation between CCNA2 and immune-cell infiltration in colon cancer. Results Database and immunohistochemical analyses indicated that CCNA2 was expressed at a significantly higher level in colon cancer tissues than normal tissues (P<0.001). The overall survival, disease-specific survival, and progression-free interval were all longer in the group with high CCNA2 expression than the group with low expression (all P<0.05). In tumor tissues, the expression level of CCNA2 decreased with increased pathological and TNM stages (P<0.05). The expression level of CCNA2 in normal tissues was consistently lower than that in colon cancer tissues across all clinical stages (all P<0.001). GSEA suggested that Wnt/β-catenin, KRAS, and other signaling pathways were enriched when CCNA2 was lowly expressed. CIBERSORT analysis revealed an increase in the infiltration of immune cells such as regulatory T cells and macrophages M0 when CCNA2 expression was low. Conclusion CCNA2 is highly expressed in colon cancer and closely associated with grade of pathology and TNM stage. It may recruit regulatory T cells through the KRAS and Wnt/β-catenin pathways, thereby reducing immune-cell infiltration and promoting colon cancer progression, leading to poor prognosis.

AIM: To investigate the molecular mechanism of microRNA-21 (miR-21) in the regulation of Schwann cell proliferation following nerve injury.METHODS:The expression of miR-2l was detected by real-time PCR. Synthetic miR-21 mimic and its control were transfected into rat Schwann cells.CCK-8 assay was performed to evaluate the influence of miR-21 on the proliferation of Schwann cells.The cell cycle distribution was determined by flow cytometry. The expression of transforming growth factorβ-induced protein ( TGFBI) and cyclin D1 were detected by Western blotting. RESULTS:The expression of miR-21 in model group was 7.87 ±0.75 and 7.75 ±0.80 times higher than that in sham operation group and blank group respectively.After transfected with miR-21 mimic, the expression of miR-21 in experimen-tal group was 2.21 ±0.14 and 2.29 ±0.21 times higher than that in negative control group and blank group respectively. Moreover, the A450 value of CCK-8 assay in experimental group at 48 h was higher than that in negative control group and blank group.The proliferation index in experimental group was higher than that in negative control group and blank group. At the same time, the expression of TGFBI obviously decreased and the cyclin D1 increased in the Schwann cells 48 h after transfection with miR-21.CONCLUSION:miR-21 promotes the proliferation activity of Schwann cells by down-regulating TGFBI expression.

This study was purpose to examine the effect of dimethyl sulfoxide (DMSO) and Tween 80 on the growth and viability of stromal cells (BMSC), colony-forming units for granulocytes and macrophages (CFU-GM) and bone marrow endothelial cell line (BMEC) from murine bone marrow in vitro, and to analyze the concentration-effect relationship. The colony yields of colony-forming units fibroblastic (CFU-F) and CFU-GM were assessed in the murine bone marrow cell cultures at various concentrations of DMSO or Tween 80 and in the control groups. The MTT assay and trypan blue exclusion were used to determine the cell viability and percentage of survival in BMSC and BMEC cultures with or without either of these organic solvents. The results showed that the colony yields of both CFU-F and CFU-GM were decreased significantly (p<0.05 or <0.01) at the concentrations (v/v final) of 2% DMSO or 0.005%-0.01% Tween 80 respectively, as compared with control. The cell viability and percentage of survival of BMSC and BMEC cultures were significantly reduced (p<0.05 or <0.01) at 0.5%-1.0% DMSO or 0.002%-0.005% Tween 80, as compared with control. With the increase of volume fractions of these solvents, the decreased percentages of corresponding measurements were increased by degrees. It is concluded that when the concentration of DMSO or Tween 80 goes to a certain level in cell culture medium, either of the organic solvents has an inhibitory action or/and cytotoxicity on the growth and viability of BMSCs, CFU-GM and BMECs. The growth inhibition and cytotoxic response are more significant at higher concentrations of these solvents.
Animals ; Bone Marrow Cells ; cytology ; Cell Line ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Cells, Cultured ; Dimethyl Sulfoxide ; pharmacology ; Dose-Response Relationship, Drug ; Endothelial Cells ; cytology ; Female ; Granulocyte-Macrophage Progenitor Cells ; cytology ; Male ; Mice ; Polysorbates ; pharmacology ; Solvents ; pharmacology ; Stromal Cells ; cytology

OBJECTIVETo observe the therapeutic effect of traditional Chinese medicine (TCM) and Western medicine (WM) treatments on mycoplasmal pneumonia in children and the changes in the serum cytokines.

METHODSNinety children with mycoplasmal pneumonia were randomly divided into the treatment group and the control group. TCM was given orally and azithromycin at the daily dose of 10 mg/kg was administered intravenously in the treatment group. In the control group, only intravenous azithromycin was given. After a 7-day treatment, the response rate, time of symptom disappearance, and serum levels of IL-6, IL-8 and TNF-alpha were observed.

RESULTSThe total response rate was 93.33% in the treatment group and 73.33% in the control group, showing a significantly better therapeutic effect in the treatment group (P<0.05). The combined treatments also showed better effects in alleviating fever, coughing and rales (P<0.05), and resulted in more obvious reduction in the serum levels of cytokines (P<0.05).

CONCLUSIONCombined treatment with TCM and WM produce good therapeutic effects in children with mycoplasmal pneumonia.

Azithromycin ; therapeutic use ; Child ; Child, Preschool ; Cytokines ; blood ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Infant ; Male ; Phytotherapy ; Pneumonia, Mycoplasma ; blood ; drug therapy ; Treatment Outcome

OBJECTIVE: To observe the effect of acupoint thread-embedding on tight junction of intestinal mucosal epithelial barrier in rats with ulcerative colitis (UC) under the state of "deficiency and stasis", and to explore its mechanism. METHODS: Sixty male SD rats were randomly divided into a control group ( RESULTS: Compared with the control group, in the model group the body weight was decreased ( CONCLUSION The thread-embedding could repair the tight junction of intestinal mucosa epithelium and reduce the permeability of intestinal mucosa epithelium, which may be related to the decrease of the expression of CaMKⅡ, MLCK and other protein kinases.
Acupuncture Points ; Animals ; Colitis, Ulcerative/therapy* ; Epithelium ; Intestinal Mucosa ; Male ; Rats ; Rats, Sprague-Dawley ; Tight Junctions

Objective To identify the chemical constituents of Xiakucao Xiaoliu mixture by high performance liquid chromatography-high resolution time-of-flight mass spectrometry (HPLC-TOF/MS). Methods The chromatographic separation ACE (3.0mm×150 mm) column was used. The mobile phase was methanol (A) and 0.1% formic acid (B). The gradient elution was: 0-5 min, 5% A; 5-10 min, 5%-15% A; 10-30 min, 15%-45%A; 30-40 min, 45%-70%B; 40-50 min, 70%-90%B. The injection volume was 2 μl. The flow rate was 0.4 ml/min. The column temperature was 25°C. The mass spectrometry was characterized by time-of-flight mass spectrometry, using ESI ion source. The common monitoring was in positive and negative ion mode. The reference ion was m/z 121.9856, 1033.9881. The scanning range was m/z 100-1200. Results A total of 37 chemical constituents were identified in the Xiakucao Xiaoliu mixture, 8 in the positive ion mode fragment voltage of 160 V, 28 in the negative ion mode fragment voltage of 160 V, and 19 in the fragment voltage of 260 V. Both positive and negative ions had 4 responses. The negative ion mode has 16 responses under both fragment voltages. And the ingredients were medicinal. Conclusion An effective method for the identification of the chemical constituents of Prunella vulgaris L. by HPLC-TOF/MS was established, which laid a foundation for its quality control and in-depth study in vivo.

Objective To determine the serum levels of tumor necrosis factor (TNF)-α,interleukin (IL)-17,IL-22 and IL-17F in patients with palmoplantar pustulosis (PP),and to estimate their relationship with disease activity in PP.Methods Venous blood samples were collected from 30 patients with PP at both active stage and stationary stage and from 20 healthy human controls.Enzyme-linked immunosorbent assay (ELISA) was conducted to determine the serum levels of TNF-α,IL-17,IL-22 and IL-17F.The paired Wilcoxon signed rank test was carried out to compare the serum levels of cytokines between patients at active stage and at stationary stage,and the Mann-Whitney U test to compare those among different groups.Results The median serum levels of TNF-α,IL-17 and IL-22 in patients with PP at active stage were 186.35 (range,113.48-412.69) ng/L,420.45 (range,278.55-748.73) ng/L and 106.48 (range,69.13-251.86) ng/L respectively,significantly higher than those at stationary stage (42.52(18.83-95.37) ng/L,48.11 (36.43-80.04) ng/L,20.32 (10.55-48.75) ng/L,respectively,all P ＜ 0.05) and those in the controls (24.30 (12.0-61.56) ng/L,10.49 (6.24-24.44) ng/L,2.58 (1.41-5.78) ng/L,respectively,all P ＜ 0.05).Moreover,the patients at stationary stage showed a significant elevation in serum levels of TNF-α,IL-17 and IL-22 compared with the controls (u =2.71,3.53,2.18,respectively,all P ＜ 0.05).No statistical difference was noted in the serum level of IL-17F among the patients at different stages and controls (P ＞ 0.05).Conclusion The circulating levels of TNF-α,IL-17 and IL-22 were associated with disease activity in PP,hinting that they may be involved in the development of PP.

Objective To investigate the therapeutic effect of Xiakucao Xiaoliu mixture on Lewis lung cancer mice. Methods 30 mice with C₅₇BL/6 mouse Lewis lung cancer xenograft model were randomly divided into three groups: model control group, Xiakucao Xiaoliu mixture group (M group), cisplatin group (DDP group). M group and DDP group were administered continuously for 14 days. Through the general observation of Lewis lung cancer mice, tumor size was determined, HE staining method was used to determine the histopathological changes of tumors, and the expression of CyclinD1 and P16 in tumor tissues was determined by immunohistochemistry. Results The tumor weight of the model control group was the heaviest, and the difference was statistically significant compared with other groups. (P<0.05). Survival state and quality of life of mice had been improved to some extent in M group. The results of tumor growth curve and HE staining in each group of mice showed that the growth of tumor cells had been inhibited and normal cells had been protected. The positive expression of CyclinD1 was significantly decreased in M group and DDP group (P<0.01), but the effect of M group on the improvement of P16 positive expression was not significant. Conclusion Xiakucao Xiaoliu mixture had a good effect on inhibiting lung tumor growth.

To study the effect of alpha1-acid glycoprotein 1 (ORM1) polymorphism on the concentration of free nortriptyline in serum, genotyping analysis was employed in ORM1 by sequencing. Eighteen unrelated male adults were chosen and given a single dose of 25 mg nortriptyline orally, then the blood samples were taken at 0, 1, 2, 3, 4, 6, 8, 12, 24, 32, 48, 72, 96 and 168 hours after drug administration. Nortriptyline and 10-OH-nortriptyline in serum and ultrafiltrate were detected for the total and free concentration by using HPLC-MS/MS. Pharmacokinetic parameters were compared among different ORM1 genotypes. No significant differences were shown in the pharmacokinetic parameters of total nortriptyline and 10-OH-nortriptyline. The mean AUC(0-infinity) of free nortritpyline in ORM1 * F/ * F1 subjects was significantly higher than that in ORM1 * F1/ * S and ORM1 * S/ * S subjects [(119.1 +/- 74.4) ng x mL(-1) x h vs (51.4 +/- 23.2) ng x mL(-1) x h and (42.4 +/- 11.6) ng x mL(-1) x h]. The percentage of protein binding in subjects with ORM1 * F1/ * F1 genotype at 2, 3, 4, 6, 8 and 12 h after administration was slightly lower than in those with ORM1 * F1/ * S and ORM1 * S/ * S genotypes while the distinct difference was shown at 4 h (P < 0.05). Different ORM1 genotypes might affect the protein binding percentage and the concentration of serum free nortriptyline. The ability binding to the drug was higher in subjects with ORM1 * S/ * S genotype than in those with other two genotypes, so as to cause the lower concentration of free nortriptyline.
Adult ; Area Under Curve ; Genotype ; Humans ; Male ; Nortriptyline ; analogs & derivatives ; blood ; pharmacokinetics ; Orosomucoid ; genetics ; metabolism ; Polymorphism, Genetic ; Protein Binding

To obtain higher potency and specificity, a series of 7-alkoxy analogues of illudalic acid was synthesized on the base of structure-activity relationship (SAR). All of these compounds exhibited submicromolar inhibition of the enzyme when tested against human leukocyte common antigen-related phosphatase (LAR) (for example, for 15e, IC50 = 180 nmol x L(-1)). They represent the most potent small-molecule inhibitors of LAR so far. These analogues also display excellent selectivity for LAR over other protein tyrosine phosphatases (PTPs) except for the highly homologous PTPsigma. The compound 15f is of 120-fold selectivity for LAR versus PTP-1B inhibition. The development of potent enzyme-specific inhibitors is so important that they may serve both as tools to study the role of LAR and as therapeutic agents for treatment of type II diabetes.