1.Cholecystokinin and nerve repair
Xuanhuang CHEN ; Libin HU ; Rongyi LI ; Hanhua CAI
Chinese Journal of Tissue Engineering Research 2013;(41):7323-7328
BACKGROUND:In the past 20 years, cholecystokinin in clinical application and nerve repair has been extensively studied. OBJECTIVE:To explore the role of cholecystokinin in nerve repair and its possible mechanism of action. METHODS:Relevant research results were retrospectively analyzed at the celland organ levels through retrieving recent literatures concerning the biological characteristics of cholecystokinin and its biological role in the nervous system. Then, we summarized the effect of cholecystokinin after nerve injury and its possible RESULTS AND CONCLUSION:Cholecystokinin and its receptors are widely distributed in the body, and under physiological and pathological conditions, their roles were complex and diverse. However, studies addressing the neuroprotective effect of cholecystokinin are not sufficient, most of which are limited to phenomenon observation. Neuroprotective mechanism of cholecystokinin is stil worthy of further studies, which can provide the basis for the clinical application.
2.Cell Apoptosis and mRNA Expression of c-fos and BNIPl in Cutaneous Squamous and Basal Cell Carcinomas
Jianqiang SHI ; Rongyi CHEN ; Yiming FAN ; Zhihua WU
Chinese Journal of Dermatology 2003;0(10):-
Objective To study the relationship between cell apoptosis and mRNA expression of c-fos and BNIPl in cutaneous squamous cell carcinoma (SCC) and basal cell carcinoma (BCC). Methods Cell apoptosis and mRNA expression of c-fos and BNIP1 in 48 SCCs and 41 BCCs were determined by TUNEL and in situ hybridization, respectively. Results Apoptosis index (AI) and c-fos mRNA expression in SCCs were higher than those in BCCs (P 0.05). AI was significantly higher in well-differentiated SCCs than in poorly differentiated SCCs (P
3.Effect of Sinomenine on Immune Function of Adjuvant Arthritis Rats
Fangjun CHEN ; Li YE ; Wei HU ; Rongyi ZHANG ; Yuanhua CHENG
Journal of Guangzhou University of Traditional Chinese Medicine 2004;0(05):-
Objective To observe the effect of sinomenine(SINO) on immune function of adjuvant arthritis(AA) rats.Methods SD rats were randomized into normal group,model group,SINO groups(treated with gastric gavage of SINO at the doses of 60,120 and 240 mg/kg respectively),and glucosides of Tripterygium Wilfordii(30 mg/kg) group.Except the normal group,the rats in other groups received subcutaneous injection of the complete Freund's adjuvant 0.1mL into the left hind foot to induce AA.The medication began from 12 days after the modeling and lasted 12 days.The pedal swelling and joint function scores were observed in different time.Radioimmunoassay was used to detect the contents of interleukin-1?(IL-1?) and tumor necrosis factor ?(TNF-?) in synovial cells.Expression of IL-1? and TNF-? mRNA was examined by reverse transcription-polymerase chain reaction(RT-PCR) assay.Results SINO at different concentrations decreased the pedal swelling and arthritis scores to various degrees,inhibited the production of IL-1? and TNF-? in the synovial cells,reduced the expression of IL-1? and TNF-? mRNA,and recovered the normal histological features of synovial cells in AA rats.Conclusion The therapeutic mechanism of SINO for rheumatoid arthritis may be related with the inhibition of secretion of inflammatory mediators in synovial cells,and with the recovery of histological features of synovial cells.
4.pEGFC1-IGFBP7 induces apoptosis of malignant melanoma cell line SK-MEL-28
Rongyi CHEN ; Yiming FAN ; Yating TU ; Hongxiang CHEN ; Qin SHEN ; Li XU
Chinese Journal of Cancer Biotherapy 2010;17(1):36-39
Objective: To construct the insulin-like growth factor binding protein 7 (IGFBP7) expression plasmid (pEGFC1-IGFBP7) and to investigate the effect of IGFBP7 on the apoptosis of SK-MEL-28 (human malignant melanoma cell line) cells. Methods: The pEGFC1-IGFBP7 plasmid was constructed; pEGFC1-IGFBP7 and empty plasmids were transfected into SK-MEL-28 cells separately. The transfection efficiency was observed under fluorescence microscope. Apoptosis of SK-MEL-28 cells after transfection was detected by Annexin-FITC/PI staining. Results: The pEGFC1-IGFBP7 plasmid was successfully constructed and was effectively transfected into SK-MEL-28 cells by Effectene reagent, with the transfection rate being 61%. The results of flow cytometry showed that pEGFC1-IGFBP7 significantly induced apoptosis of SK-MEL-28 cells, with the apoptotic rates of pEGFC1-IGFBP7, empty vector, and non-transfected plasmid groups being (28.4±2.57)%, (5.8±0.44)%, and (6.4±0.71)% 24 h after transfection, respectively (F=406.138, P<0.05). Conclusion: pEGFC1-IGFBP7 can effectively induce apoptosis of malignant melanoma SK-MEL-28 cells, which provides an experimental basis for IGFBP7 gene-based therapy of malignant melanoma.
5.Effects of progesterone on PMN-mediated inflammatory responses to gonococcal infection
Jun SHUAI ; Hongxiang CHEN ; Zhihong WU ; Rongyi CHEN ; Li XU ; Ying YU ; Yating TU
Chinese Journal of Dermatology 2010;43(5):324-327
Objective To evaluate the effects of progesterone on polymorphonuclear leukocyte (PMN)-mediated inflammatory responses to gonococcal infection. Methods Peripheral neutrophils were isolated from heparinized peripheral blood obtained from normal individuals, then divided into 4 groups: progesterone group (pretreated with progesterone only), gonococcus group (stimulated with gonococcal suspension), intervention group (pretreated with progesterone followed by stimuation with gonococcal suspension), and control group (receiving no pretreatment or stimulation). Real-time RT-PCR was conducted to detect the mRNA expression levels of inducible nitric oxide synthase (iNOS), tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β)in neutrophils from all groups at 0, 3, 8, 12 and 24 hours after the last treatment, and iNOS protein levels were measured by Western-blot in gonococcus group and intervention group. Results Real-time RT-PCR indicated that the expression levels of iNOS, TNF-α and IL-1β mRNA increased in gonococcus group and intervention group, and reached their peak at 8 hours in gonococcus group, while no significant changes were noted in the above parameters in progesterone group or control group. Also, the level of iNOS, TNF-α and IL-1β mRNA was lower in intervention group than that in the gonococcus group (P < 0.05). Western blot showed an elevation in iNOS protein expression in both gonococcus group and intervention group, and the former group was higher than the latter group in the parameter (P < 0.05). Conclusions Progesterone can downregulate the expressions of iNOS, TNF-α and IL-1 β by PMNs, inhibit the PMN-induced inflammatory responses induced by gonococcal infection, which is likely to be associated with the asymptomatic gonococcal infection in women.
6.Some factors correlated with the symptoms of female gonococcal infections
Zhihong WU ; Hongxiang CHEN ; Rongyi CHEN ; Xianfeng FANG ; Jan SHUAI ; Li XU ; Ying YU ; Yating TU
Chinese Journal of Dermatology 2009;42(5):311-314
Objective To study the relationship of symptoms of female gonococcal infections to Chlamydia trachomatis infection, serum sex hormone levels, etc. Methods A total of 136 gonorrhea female patients without obvious symptoms were recruited in this study together with 45 gonorrhea patients with obvious symptoms as the controls. Serum progesterone (P) and estradiol (E2) levels were measured by radio immunoassay (RIA). Cervical swabs were obtained from the subjects and eluted into isotonic saline solution, the elution was divided into 2 portions and tested for the levels of TNF-α and IL-1β by ELISA and for the DNA of C. Trachomatis and N. Gonorrhea with PCR. Statistical analysis was carried out by SPSS for Windows (version 12.0). Results There was no statistical correlation between C. Trachomatis infection and asymptomatic status of female gonococcal infection (χ2 = 0.016, P > 0.05). However, the decrease in the level of TNF-α and IL-1β significantly correlated with the increase in serum progestogen (r = -0.8798, -0.8935, respectively, both P < 0.01). Conclusion The high serum level of progesterone may be associated with the asymptomatic status of gonococcal infection.
7.The nongenomic effects of progesterone in repressing iNOS activation through P38MAPK pathways in gonococci-infected polymorphonuclear leukocytes and the clinical significance.
Rongyi, CHEN ; Yating, TU ; Jiaxi, LIN ; Weibing, SHE ; Juan, LI ; Zhihong, WU ; Li, XU ; Hongxiang, CHEN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2010;30(1):119-25
Progesterone has nongenomic effects on inducible nitric oxide synthase (iNOS), which is mediated by mitogen activated protein kinase (MAPK) pathways. This effect is supposed to have some potential association with asymptomatic gonococcal infections in women by immunological depression. In this study, polymorphonuclear leukocytes (PMNs) challenged by gonococci were used to study the nongenomic effects of progesterone. The activation of iNOS was assessed by measuring [(3)H] L-arginine converses to [(3)H] L-citrulline, and the activity of MAPK was detected by Western blot. It was found that the activity of iNOS and the yields of NO were enhanced significantly in gonococci-challenged PMNs compared with the controls (P<0.01). Progesterone could repress the activation of iNOS through P38MAPK pathway within PMNs (P<0.05), which could be blocked by SB203580 (P<0.01), but not by actinomycin D (P>0.05). It was also found subsequently that in the serum specimens collected from gonococci-infected but asymptomatic women, the progesterone level was higher than that in women with severe symptoms (P<0.01). Moreover, the yield of NO had an inverse correlation with progesterone. With these results it suggested that the rapid nongenomic effects of progesterone may inhibit iNOS activation and NO yields mediated by P38MAPK pathways, which were supposed to be concerned with asymptomatic women infected with gonococci.
8.Possible mechanisms of cholecystokinin promoting sciatic nerve regeneration
Xuanhuang CHEN ; Rongyi LI ; Guodong ZHANG ; Haibin LIN ; Xianwei WU ; Yujin LIN ; Feng ZHENG
Chinese Journal of Tissue Engineering Research 2014;(11):1700-1705
BACKGROUND:Previous studies have found that cholecystokinin octapeptide (CCK-8) can promote the regeneration after sciatic nerve injury in rats, but the exact mechanism remains unclear.
OBJECTIVE:To screen effective indicators and analyze the mechanism of CCK-8 promoting sciatic nerve regeneration from the perspective of nerve growth factor and nerve regeneration microenvironment.
METHODS:Healthy Sprague-Dawley rats, for the preparation of unilateral sciatic nerve transection injury model, were randomly divided into two groups. In the CCK-8 group, the animal model received intraperitoneal injection of CCK-8 (8 nmol/kg) for consecutive 7 days, while the control group was injected with equal volume of normal saline. The nerve growth factor expression, inducible nitric oxide synthase in the spinal cord, serum superoxide dismutase activity and malondialdehyde concentration, as wel as apoptotic cel s in spinal cord were al detected.
RESULTS AND CONCLUSION:In the CCK-8 group, nerve growth factor expression was higher than that in the control group (P<0.01), while inducible nitric oxide synthase and the number of apoptotic cel s were lower (P<0.01), serum superoxide dismutase activity was higher but malondialdehyde concentrations was lower (P<0.01, 0.05). The mechanisms of CCK-8 promoting sciatic nerve regeneration include protecting neurons, anti-apoptosis, inhibiting inflammatory response, anti-NO and anti-oxidation, reducing malondialdehyde, and al eviating free radical damage, as wel as stimulating nerve growth factor expression and release.
9.Expression of retinoic acid-related orphan receptors and interleukin-17 in mice with vaginal candidiasis
Dan LUO ; Jin′e ZHANG ; Rongyi CHEN ; Yanping YANG ; Ying ZHOU ; Yiming FAN ;
Chinese Journal of Dermatology 2017;50(1):33-38
Objective To investigate the role of T helper 17 cells/interleukin?17(Th17/IL?17) axis in the occurrence of vaginal candidiasis in mice. Methods A total of 120 female BALB/c mice were randomly and equally divided into Ei, En, Ci and Cn groups. Three days before vaginal inoculation, estrogen (Ei and En)groups and control(Ci and Cn)groups received subcutaneous injection of 0.05 mg estradiol and 0.1 ml sterilized soybean oil at the hind legs, respectively, and then the hormone treatment continued every other day until the end of experiment. Infected(Ei and Ci)groups and noninfected(En and Cn) groups were inoculated intravaginally with 10μl(5 × 104 conidia)of Candida albicans suspension and 10μl of sterilized phosphate?buffered saline, respectively. Ten mice were randomly selected from each group and sacrificed on day 3, 7 and 14 after inoculation. The intact vagina tissues were resected and then frozen in liquid nitrogen or embedded in paraffin. Real?time fluorescence?based quantitative PCR(qRT?PCR)and immunofluorescent staining were performed to measure mRNA expression and immunofluorescence intensities of retinoic acid?related orphan receptorγt(RORγt), RORα and IL?17, respectively. Western blot analysis was conducted to determine protein expression of RORγt and IL?17. Results Laser scanning confocal microscopy showed that RORγt, RORα and IL?17 immunofluorescence was mainly located at inflammatory cells of the lamina propria and blood vessels in En and Cn groups, at mucosal epithelium, adherent hyphae, and inflammatory cells of the lamina propria and blood vessels in Ci group, and at mucosal epithelium, vaginal canal and endocytosed hyphae, and inflammatory cells of the lamina propria and blood vessels in Ei group. qRT?PCR and immunofluorescent staining uncovered that mRNA expression and immunofluorescence intensities of RORγt, RORα and IL?17 were significantly higher in En, Ci and Ei groups than in Cn group at the same time points(all P<0.05), as well as in the Ei group than in En and Ci groups(both P<0.05), and were increased gradually over time in En, Ci and Ei groups, but not in the Cn group. Additionally, mRNA expression and immunofluorescence intensities of RORγt and RORαand IL?17 generally peaked on day 14 after inoculation, while the immunofluorescence intensity of IL?17 peaked on day 7 (P < 0.05). Western blot analysis revealed that protein expression of RORγt and IL?17 was significantly higher in the infected(Ei and Ci)groups than in the noninfected(En and Cn)groups at the same time points(RORγt:F=45.685, P<0.001;IL?17:F=29.655, P<0.01), and was highest in the Ei group(P<0.05);however, no significant differences were observed between Cn and En groups(both P>0.05). Moreover, RORγt and IL?17 protein expression in Ci and Ei groups was obviously up?regulated on day 7 after inoculation (RORγt: F = 13.137, P < 0.001; IL?17: F = 11.182, P < 0.001), but was not increased further on day 14. Conclusion Vaginal candida infection can up?regulate the expression of RORγt, RORα and IL?17, suggesting that Th17/IL?17 axis may be involved in the occurrence of vaginal candidiasis in BALB/c mice.
10.Plumbagin enhances TRAIL-mediated apoptosis through up-regulation of death receptor in human melanoma A375 cells.
Jiawen, LI ; Qin, SHEN ; Rui, PENG ; Rongyi, CHEN ; Ping, JIANG ; Yanqiu, LI ; Li, ZHANG ; Jingjing, LU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2010;30(4):458-63
Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a promising anti-cancer agent. However, emergence of drug resistance limits its potential use. Plumbagin is a natural quinonoid compound isolated from plant. In this study, induced apoptosis effect of the combined treatment with plumbagin and TRAIL on human melanoma A375 cell line was examined and possible mechanism was investigated. The cells were divided into four groups: control group, plumbagin group (plumbagin, 5 or 10 mumol/L), TRAIL group (TRAIL, 30 ng/mL) and plumbagin+TRAIL group (combined treatment group). The apoptosis, and the expression of DR4 and DR5 were detected by flow cytometry. The activities of caspase-8 and caspase-3 were determined by colorimetric assay. The results showed that the apoptosis rate was 8.3% in TRAIL group, 10.35%-16.94% in plumbagin group and 52.39%-65.39% in combined treatment group, respectively, with the difference being significant between combined treatment group and plumbagin or TRAIL group (P<0.05 for each). Moreover, plumbagin alone could markedly up-regulate DR5 mRNA and protein expression, and slightly increase DR4 mRNA and protein expression. Treatment of human melanoma A375 cells with plumbagin resulted in the activation of Caspase-3, but not Caspase-8. These results suggest that plumbagin might be useful for TRAIL-based treatment for melanoma.