1.Xinfukang drug-containing serum on bone marrow stem cell transcription and secretion of SDF-1α
Bohong KAN ; Yijing WANG ; Xiantong HU ; Qing GAO ; Yingchang FAN ; Yingna HE ; Kongyin ZHAO
International Journal of Biomedical Engineering 2014;37(5):275-278
Objective To clarify the effects of Xinfukang containing-serum on stromal cell-derived factor-1α (SDF-1α) translation and protein secretion of bone marrow stem cells (BMSCs).Methods BMSCs were isolated and amplified using bone marrow culture method,and were identified by flow cytometry.mRNA and protein secretion of SDF-1α were detected by quantitative PCR (q-PCR) and enzyme linked immunosorbent assay (ELISA),respectively.Results The expression of SDF-1α mRNA were significantly increased after 72 h in drug-containing serum,and SDF-1α mRNA in the experimental group was approximately 200 times as that in the control group (P<0.05).Secretion of SDF-1 α in the experimental group (277.561 1 ± 15.651 8) pg/ml was nearly doubled compared with that in the control group (153.107 1±14.765 1) pg/ml (P<0.05).Conclusions BMSCs from whole bone marrow adherent culture have high purity,and drug-containing serum can promote BMSCs to express SDF-1 α mRNA and secretion of SDF-1 α.
2.Establishment of rat skin organ culture model and initiation of hair follicular morphogenesis with its correlative dynamic expression of Wnt10 b/β-catenin
Yingchang JI ; Yu LI ; Feng LU ; Zhiqi HU ; Sen WANG ; Changmin LIN ; Jianhua GAO
Chinese Journal of Medical Aesthetics and Cosmetology 2010;16(2):116-120
Objective To establish a rat skin organ culture model to study the initiation of hair follicle morphogenesis, and the dynamic expression of Wnt10b/β-catenin in the developing hair follicle. Methods The dorsal skins of SD rat at embryos 14-17 (E14-E17) were cultured on a gelatin sponge-supported tissue culture system at the air/liquid interface of DMEM with 10 % fetal bovine serum (FBS) for 3-6 days, some of which removed from El5 were cultured in DMEM with FBS of different concentrations. HE staining and fluoroimmunoassay were adopted. Results The model we established allowed skin tissues isolated from E14-E15 to develop in a manner that was histologically and temporarily similar to the process in vivo. However, the developing hair follicle ceased to continue when their morphogenetic process reached the forth stage, and the concentration of FBS did not show any significant effect on the development of hair follicle. Expression of Wnt10b/β-catenin was induced in culture, as it also occurred in vivo,but grew weak till it disappeared in culture for 6 days, which was accompanied by development halt of hair follicle. Conclusions A rat skin organ culture model is established in which the morphogenesis of hair follicle takes place in a manner similar to in vivo, and Wnt10b/β-catenin probably has a close connection with the early morphogenesis of hair follicle.
3.Application of diffusion weighted magnetic resonance imaging in assessing the efficacy of chemoradiotherapy for uterine cervical carcinoma
Jiansheng LI ; Zhijun CHEN ; Rijie TANG ; Hainan ZHANG ; Bingui LU ; Yingchang GAO
Cancer Research and Clinic 2014;26(11):729-732,736
Objective To explore the feasibility of diffusion-weighted imaging (DWI) in assessing the efficacy of chemoradiotherapy for cervical cancer.Methods Data of magnetic resonance imaging (MRI) and DWI were analyzed in 32 patients with uterine cervical cancer received conventional prior to chemoradiotherapy and after 1 and 3 months of therapy.13 cases of normal controls also had been examed by MRI and DWL DWI with b values of (0,300) s/mm2 and b values of (0,600) s/mm2 were performed in all patients.Pretreatment post-treatmentADC values were compared between the health group and patients group.Results When the b =300 s/mm2,normal cervical average ADC value was (1.72±0.31)×10-3 mm2/s,cervical cancer was (1.10±0.24)×10-3 mm2/s before treatment and was (1.61±0.23)×10-3 mm2/s after treatment.When the b =600 s/mm2,normal cervical average ADC value was (1.46±0.25)×10-3 mm2/s cervical cancer was (0.89±0.21)×10-3 mm2/s before treatment and was (1.54±0.18)×10-3 mm2/s after treatment.When b =300 s/mm2,ADC value was higher than when b =600 s/mm2.ADC values of cervical cancer was significantly lower than that of the normal cervix group,ADC values of cervical lesions after chemoradiotherapy was significantly higher than that before chemoradiotherapy (P < 0.05).In the same group with different b values,ADC value was not significant (P > 0.05).Conclusion Joint observation of DWI and ADC values could be more objective and accurate in the analysis of the disease and would help to evaluate the efficacy of chemoradiotherapy.
4.Effects of exogenous hydrogen sulfide on intestinal function in rabbits after cardiopulmonary resuscitation
Rui GONG ; Yingchang XUE ; Hao XU ; Liufang GAO ; Guiyang LU ; Jiating ZHOU ; Minwei ZHANG
Chinese Critical Care Medicine 2018;30(12):1178-1183
Objective To investigate the effect of exogenous hydrogen sulfide (H2S) on intestinal mucosal barrier after cardiopulmonary resuscitation (CPR) in cardiac arrest (CA) rabbits. Methods Forty-four male New Zealand rabbits were divided into sham operation group (Sham group, n = 12), post-cardiac arrest syndrome (PCAS) group (n = 16) and H2S intervention group (PCAS+NaHS, n = 16) according to random number table method. The rabbit model of PCAS was established by tracheal clamping and suffocation, and CPR was started at 5 minutes after CA. However, Sham group did not clamp the tracheal intubation after anesthesia, and the other operations were the same as those in PCAS group. In the PCAS+NaHS group, a bolus of NaHS (0.5 mg/kg), a H2S donor, was injected via era vein 1 minute before the start of CPR, followed by a continuous injection of NaHS (1.5 mg·kg-1·h-1) for 3 hours, while the rabbits in other group were intravenously injected with the same volume of normal saline (NaCl 0.9%). Intestinal and portal vein blood samples were collected 24 hours after return of spontaneous circulation (ROSC). The level of serum fluorescein isothiocyanate-dextran (FD-4) was detected by fluorescein isothiocyanate (FITC) labeling method to reflect intestinal mucosal permeability. After hematoxylin-eosin (HE) staining of small intestine tissues, the morphological changes of mucosa were observed under light microscope, and the intestinal mucosa injury score was calculated. The expression of tight junction protein ZO-1 in intestinal mucosa was detected by immunohistochemistry. The content of malondialdehyde (MDA) in small intestinal tissue was determined by thiobarbituric acid chromogenic method, the activity of superoxide dismutase (SOD) was determined by xanthine oxidation method, and the level of myeloperoxidase (MPO) was determined by double antibody sandwich enzyme linked immunosorbent assay (ELISA) to reflect the oxidative stress and inflammatory reaction in small intestinal tissue. The expression of apoptosis protein (caspase-3) and autophagy related protein (Beclin-1, LC3) in small intestine tissue was detected by Western Blot. Results 12, 13 and 14 animals were successfully resuscitated in Sham group, PCAS group and PCAS+NaHS group respectively, while 12 animals in each group survived to the end of experiment. Compared with Sham group, the level of FD-4 in portal vein serum was significantly increased in PCAS group (mg/L: 11.95±0.59 vs. 1.43±0.48, P < 0.05), the pathological injury and inflammation infiltration were obviously aggravated under light microscope, the score of small intestine injury was significantly increased (4.21±0.37 vs. 0.36±0.18, P < 0.05), the expression of tight junction protein ZO-1 in the intestine was visibly down-regulated detected by immunohistochemistry, MDA content and MPO activity were significantly increased [MDA (nmol/mg): 3.65±0.32 vs. 1.54±0.24, MPO (U/g): 362±35 vs. 134±18, both P < 0.05], while SOD activity was significantly decreased (U/mg:78.84±7.49 vs. 115.48±8.48, P < 0.05), the expression levels of cleaved capase-3, Beclin-1 and LC3 proteins in the intestine were significantly increased (caspase-3/β-actin: 1.11±0.08 vs. 0.21±0.02, Beclin-1/β-actin: 2.08±0.11 vs. 0.42±0.03, LC3/β-actin: 1.05±0.07 vs. 0.37±0.05, LC3-Ⅱ/ LC3-Ⅰ: 1.28±0.14 vs. 0.17±0.02, all P < 0.05). Compared with PCAS group, the portal vein serum FD-4 level in PCAS+NAHS group was significantly decreased (mg/L:5.59±0.48 vs. 11.95±0.59, P < 0.05), the intestinal mucosal pathological injury and inflammatory cell infiltration were significantly decreased, the score of small intestine injury was significantly decreased (2.18±0.47 vs. 4.21±0.37, P <0.05), the expression of ZO-1 in intestine was significantly increased, MDA content and MPO activity in intestine were significantly decreased [MDA (nmol/mg): 2.65±0.31 vs. 3.65±0.32, MPO (U/g): 251±21 vs. 362±35, both P < 0.05], while SOD activity was significantly increased (U/mg: 96.86±7.52 vs. 78.84±7.49, P < 0.05), while the expression of activated caspase-3, Beclin-1 and LC3 proteins was significantly decreased (caspase-3/β-actin: 0.72±0.06 vs. 1.11±0.08, Beclin-1/β-actin: 0.96±0.08 vs. 2.08±0.11, LC3/β-actin: 0.72±0.06 vs. 1.05±0.07, LC3-Ⅱ/ LC3-Ⅰ:0.83±0.09 vs. 1.28±0.14, all P < 0.05). Conclusion H2S has a protective effect on intestinal mucosal injury induced by CA/CPR, which may be related to tight junction protein ZO-1 up-regulation, oxidative stress alleviation, inflammation reduction, apoptosis and autophagy inhibition.