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OBJECTIVE To study the effects and potential mechanism of wogonin （Wog） on airway inflammation in rats with chronic obstructive pulmonary disease （COPD）. METHODS Eighty-four rats were randomly divided into control group， model group， Wog low-dose and high-dose groups （intragastric administration of 50， 100 mg/kg）， aminophylline group （positive control， intragastric administration of 2.3 mg/kg）， recombinant rat receptor-interacting protein kinase 1 [rRIPK1， receptor-interacting protein kinase 1 （RIPK1） activator] group （tail vein injection of 8 µg/kg）， and Wog high-dose+rRIPK1 group （intragastric administration of Wog 100 mg/kg+tail vein injection of rRIPK 8 µg/kg）， with 12 rats in each group. Except for control group， COPD model of other groups was induced by smoking combined with tracheal injection of lipopolysaccharide. Twenty-four hours after successful modeling， the rats were administered once a day for 4 weeks. The changes of peak inspiratory flow （PIF）， peak expiratory flow （PEF） and minute ventilation （MV），forced expiratory volume in one second（FEV1）/forced vital capacity（FVC） were measured after the last medication； the serum levels of interleukin 1β（IL-1β）， IL-6 and tumor necrosis factor-α （TNF-α） were measured by ELISA； the pathological changes of lung tissue in rats were observed； the apoptotic rate of pulmonary epithelial cells was detected. mRNA expressions of RIPK1， RIPK3 and mixed lineage kinase domain-like protein （MLKL）， and protein expressions of RIPK1， RIPK3 and p-MLKL were all detected in lung tissue of rats. RESULTS Compared with control group， PIF， PEF， MV and FEV1/FVC of model group were decreased significantly （P＜0.05）， while the levels of IL-1β， IL-6 and TNF- α were increased significantly （P＜0.05）； there was a large number of inflammatory cells infiltration in the lung tissue and bronchialwall thickening in model group； the apoptotic rate of pulmonary epithelial cells，mRNA expressions of RIPK1， RIPK3 and MLKL， protein expressions of RIPK1， RIPK3 and p-MLKL were increased significantly （P＜0.05）. Compared with model group， above indexes of rats were improved significantly in Wog low-dose and high-dose groups （P＜0.05）， and pathological injuries were alleviated significantly. The corresponding indexes of rats were worsened in rRIPK1 group （P＜0.05）， and pathological damage had further worsened. rRIPK1 significantly attenuated the inhibitory effect of high-dose Wog on airway inflammation and RIPK1/RIPK3/ MLKL pathway in COPD rats （P＜0.05）. CONCLUSIONS Wog may improve airway inflammation in COPD rats by inhibiting RIPK1/RIPK3/MLKL signal pathway.
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Objective:To evaluate associations of psoriasis with adverse pregnancy outcomes.Methods:Databases, including CNKI, Wanfang, VIP, PubMed, Embase and Cohrane Library databases, were searched for published articles on associations between psoriasis and adverse pregnancy outcomes between January 1980 and December 2018. Quality of included articles was assessed by using MOOSE checklist. Statistical analysis was carried out with Review Manager 5.3 software.Results:One cohort study, 4 case-control studies and 2 cross-sectional studies meet the inclusion criteria. After heterogeneity test, meta-analysis was carried out by using a random effect model. The risks of cesarean ( OR= 1.17, 95% CI: 1.01- 1.37) , eclampsia or preeclampsia ( OR= 1.34, 95% CI: 1.00- 1.79) and premature birth ( OR= 1.09, 95% CI: 1.00- 1.19) were significantly higher in patients with psoriasis than in individuals without psoriasis (all P < 0.05) . There was no significant difference between patients with psoriasis and individuals without psoriasis in the risks of spontaneous abortion, low birth weight, high birth weight, stillbirth, congenital malformation, placental abruption, overdue delivery, low Apgar score (< 7) , polyhydramnios and oligohydramnios. Conclusion:The risks of cesarean, eclampsia or preeclampsia, and premature birth are higher in patients with psoriasis than in individuals without psoriasis.
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Objective@#To evaluate the effect of epigallocatechin gallate (EGCG) on T helper cell 1 (Th1) and Th2 in psoriasis patients.@*Methods@#A total of 33 patients with plaque-type psoriasis vulgaris were enrolled, and peripheral blood mononuclear cells (PBMC) were isolated and cultured. The appropriate concentration of EGCG was determined by methyl thiazol tetrazolium (MTT) assay. PBMC at exponential growth phase were divided into 2 groups to be treated with EGCG (EGCG group) or not (control group) for 24 hours. Flow cytometry was performed to determine proportions of Th1 and Th2 cells, enzyme-linked immunosorbent assay (ELISA) to detect levels of Th1 (interleukin[IL]-2, interferon[IFN]-γ) and Th2 cytokines (IL-4, IL-10) in the cell culture supernatant, and real-time quantitative RCR (qRT-PCR) to determine the mRNA expression of T-bet (a Th1 transcription factor) and GATA3 (a Th2 transcription factor) . Statistical analysis was carried out by using t test.@*Results@#According to the MTT assay results, EGCG at a non-toxic concentration of 60 μmol/L was chosen for subsequent experiments. Compared with the control group, the EGCG group showed significantly decreased number of Th1 cells (t = 3.43, P = 0.026) , increased number of Th2 cells (t = 6.68, P = 0.026) , and decreased Th1/Th2 ratio (P < 0.05) . The levels of IL-2 and IFN-γ in the culture supernatant of PBMC were both significantly lower in the EGCG group (824.45 ± 101.21 ng/L, 1 623.62 ± 185.56 ng/L respectively) than in the control group (1 568.32 ± 196.45 ng/L, 3 287.63 ± 235.54 ng/L respectively) , while the levels of IL-4 and IL-10 were significantly higher in the EGCG group (389.48 ± 46.63 ng/L, 285.95 ± 53.28 ng/L respectively) than in the control group (225.38 ± 26.92 ng/L, 165.46 ± 32.25 ng/L respectively) . Compared with the control group, the EGCG group showed significantly decreased T-bet mRNA expression (t = 11.99, P < 0.001) , but increased GATA3 mRNA expression (t = 18.62, P < 0.001) .@*Conclusion@#EGCG can reduce the number of Th1 cells, inhibit the production of Th1 cytokines and transcription factors, and increase the number of Th2 cells and the production of Th2 cytokines and transcription factors, followed by the modulation of Th1/Th2 immune imbalance.
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Objective To evaluate the effect of epigallocatechin gallate (EGCG) on T helper cell 1 (Th1) and Th2 in psoriasis patients.Methods A total of 33 patients with plaque-type psoriasis vulgaris were enrolled,and peripheral blood mononuclear cells (PBMC) were isolated and cultured.The appropriate concentration of EGCG was determined by methyl thiazol tetrazolium (MTT) assay.PBMC at exponential growth phase were divided into 2 groups to be treated with EGCG (EGCG group) or not (control group) for 24 hours.Flow cytometry was performed to determine proportions of Th 1 and Th2 cells,enzyme-linked immunosorbent assay (ELISA) to detect levels of Th1 (interleukin [IL]-2,interferon [IFN]-γ) and Th2 cytokines (IL-4,IL-10) in the cell culture supernatant,and real-time quantitative RCR (qRT-PCR) to determine the mRNA expression of T-bet (a Th1 transcription factor) and GATA3 (a Th2 transcription factor).Statistical analysis was carried out by using t test.Results According to the MTT assay results,EGCG at a non-toxic concentration of 60 μmol/L was chosen for subsequent experiments.Compared with the control group,the EGCG group showed significantly decreased number of Th1 cells (t =3.43,P =0.026),increased number of Th2 cells (t =6.68,P =0.026),and decreased Th1/Th2 ratio (P ＜ 0.05).The levels of IL-2 and IFN-γin the culture supernatant of PBMC were both significantly lower in the EGCG group (824.45 ± 101.21 ng/L,1 623.62 ± 185.56 ng/L respectively) than in the control group (1 568.32 ±196.45 ng/L,3 287.63 ± 235.54 ng/L respectively),while the levels of IL-4 and IL-10 were significantly higher in the EGCG group (389.48 ± 46.63 ng/L,285.95 ± 53.28 ng/L respectively) than in the control group (225.38 ± 26.92 ng/L,165.46 ± 32.25 ng/L respectively).Compared with the control group,the EGCG group showed significantly decreased T-bet mRNA expression (t =11.99,P ＜ 0.001),but increased GATA3 mRNA expression (t =18.62,P ＜ 0.001).Conclusion EGCG can reduce the number of Th1 cells,inhibit the production of Th 1 cytokines and transcription factors,and increase the number of Th2 cells and the production of Th2 cytokines and transcription factors,followed by the modulation of Th 1/Th2 immune imbalance.
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Acupoints are the response points of diseases and the stimulated points by acupuncture and moxibustion. The authors take,a's extra point,and Hegu(LI 4) as examples to explore the changing cha-racteristics of acupoints from the aspects of finding the response points based on groping acupoints,the features of diagnosis and treatment and action mechanism. It is believed that Hegu(LI 4) is located on the radialis side of the forefinger,and in the changeable status according to the function of the body. While,a radialis point of the second metacarpal,is related to Hegu(LI 4) which is moving in pathological situation. The conclusion is that other acupoints may be bound up with the body function and in changeable status.
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Objective To evaluate effects of ursolic acid (UA) on interleukin?33 (IL?33) expression in HaCaT cells induced by interferon?γ(IFN?γ), and to explore their mechanism. Methods Some HaCaT cells were treated with UA at different concentrations(0, 0.1, 1, 5, 10, 20, 40 and 80μmol/L)for 24, 48 and 72 hours separately. Then, methyl thiazolyl tetrazolium(MTT)assay was conducted to evaluate cell proliferative activity. A cell model of inflammation was established by culture of HaCaT cells with the presence of 200μg/L IFN?γ. Some HaCaT cells were classified into several groups to be treated with IFN?γ(200μg/L)and UA(10 and 15μmol/L)alone or in combination (firstly treated with IFN?γ followed by UA treatment), and those receiving no treatment served as the blank control group. Reverse transcription PCR (RT?PCR) was performed to detect mRNA expressions of IL?6 and IL?33, and Western?blot analysis to measure IL?33 protein expression after 12?hour culture. The expressions of extracelluar signal?regulated kinase 1/2(ERK1/2)and phosphorylated ERK1/2(p?ERK1/2)were also measured by Western?blot analysis after 5?and 60?minute treatments with IFN?γand UA alone or in combination. Results MTT assay showed that the treatments with 5-20μmol/L UA for 24 hours had no effects on cell proliferative activity, while 40-80μmol/L UA could significantly inhibit it at 24, 48 and 72 hours (all P 0.05), while IL?33 protein expression was significantly lower in the IFN?γ+15?μmol/L UA group than in the IFN?γ+10?μmol/L UA group(P<0.05). The p?ERK1/2 protein expression significantly increased in HaCaT cells treated with IFN?γ for 5 and 60 minutes compared with the blank control group, but significantly decreased in the IFN?γ+15?μmol/L UA group compared with the IFN?γgroup(0.458 ± 0.053 vs. 0.941 ± 0.042 at 5 minutes, 0.302 ± 0.054 vs. 0.509 ± 0.032 at 60 minutes, both P < 0.05). However, no significant differences were observed in the total ERK1/2 protein expression between the IFN?γ+15?μmol/L UA group and IFN?γgroup at 5 or 60 minutes. Conclusion UA can suppress IL?33 expression in HaCaT cells induced by IFN?γ, likely by regulating expressions of the ERK signaling pathway?related proteins.
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Objective To evaluate the protective effect of epigallocatechin gallate (EGCG) against interleukin (IL)-17-induced injury to keratinocytes,and to explore its mechanism.Methods Some cultured HaCaT cells were divided into 3 groups to be treated with IL-17 alone at concentrations of 50,70,90 μg/L,respectively,with those receiving no treatment as the blank control group.Some HaCaT cells were divided into 5 groups:IL-17 group treated with 90 μg/L IL-17 alone,IL-17 + EGCG group treated with 90 μg/L IL-17 and 60 μmol/L EGCG,IL-17 + SP600125 group treated with 90 μg/L IL-17 and SP600125 (a JAK signaling pathway inhibitor),IL-17+ EGCG + anisomycin group treated with 90μg/L IL-17,60xmol/L EGCG and anisomycin (a Janus kinase signaling pathway activator),and blank control group receiving no treatment.After different durations of treatment,CCK-8 assay was performed to evaluate cellular proliferative activity,flow cytometry to detect cell apoptosis,enzyme-linked immunosorbent assay (ELISA) to measure expression levels of IL-6,IL-23 and IL-8,and Western-blot analysis to determine protein expressions of c-Jun N-terminal kinase (JNK) and phosphorylated JNK (P-JNK).Results IL-17 promoted cellular proliferation of HaCaT cells,and the proliferation rate,which was correlated with the concentration of IL-17,reached the maximum in the 90-μg/L IL-17 group (P ＜ 0.05).EGCG at 60 μmol/L significantly inhibited cellular proliferation of,promoted apoptosis in,and reduced IL-6,IL-23 and IL-8 expressions in,HaCaT cells induced by 90 μg/L IL-17 (all P ＜ 0.05).Compared with the IL-17 group,the IL-17 + EGCG group and IL-17 + SP600125 group both showed significantly decreased P-JNK expression,cell proliferation rate and IL-6,IL-23 and IL-8 expression levels (all P ＜ 0.05).However,compared with the IL-17 + EGCG group,the IL-17 + EGCG + anisomycin group showed significantly increased protein expression of P-JNK,cell proliferation rate and IL-6,IL-23 and IL-8 expression levels (all P ＜ 0.05).Conclusion EGCG protected against IL-17-induced injury to HaCaT cells,such as abnormal cell proliferation,apoptosis and inflammatory response,likely by inhibiting the JNK signaling pathway.
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Objective] To sum up director doctor He Ruoping’s by stages treatment experience in treating bladder cancer. [Method]By learning from director doctor He Ruoping for many years and combining my own clinical experience, from the etiology and pathogenesis, therapeutic principle of TCM syndrome differentiation treatment characteristics, etc, it sums up the teacher He ’s unique feature of by stages treatment of bladder cancer, and with 1 case for detailed explanation. [Result]Teacher He advocating the combination of traditional Chinese medicine and western medicine, puts forward the principle of by stages treatment to bladder cancer:treatment of postoperative perfusion stage, treatment of following up stage without perfusion and treatment of palliative transfer stage, and makes prescriptions in line with the characteristics of each stage. [Conclusion] Teacher He ’s by stages treatment to bladder cancer obtains the good curative effect, having enlightenment meaning to clinical practice, with promotion value.
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Objective To investigate the effect of downregulation of KIAA0101 protein expression on the proliferation and invasion of a cutaneous squamous cell carcinoma cell line SCL-1,and to explore possible molecular mechanisms underlying the effect.Methods SCL-1 cells were classified into three groups: siRNA control group transfected with the control siRNA,KIAA0101 group transfected with KIAA0101 siRNA,and untreated group remaining untreated.After additional culture,Western blot was used to detect the expression of KIAA0101 protein and proteins associated with cell proliferation and invasion,cell counting kit-8 (CCK-8) to evaluate cellular proliferative activity,and Boyden chamber assay to estimate invasive ability of cells.Results The relative expression level of KIAA0101 protein was 0.062 ± 0.095 in the KIAA0101 group,significantly lower than that in the untreated group (0.359 ± 0.044,P ＜0.05) and siRNA control group (0.379 ± 0.025,P ＜0.05).A significant decrease was observed in cellular proliferative activity (from 24 to 96 hours) and invasive activity (at 48 hours) in the KIAA0101 group compared with the other two groups (all P ＜0.05).Moreover,compared with the untreated group and siRNA control group,the KIAA0101 group showed a stronger expression of p21 protein (0.570 ± 0.060 vs.0.048 ± 0.018 and 0.055 ± 0.014,P ＜0.01) but a weaker expression of matrix metalloproteinase 2 (MMP2) protein (0.051 ± 0.013 vs.0.205 ± 0.029 and 0.221 ± 0.029,P ＜0.01).Conclusion The inhibition of SCL-1 cell proliferation and invasion induced by the downregulation of KIAA0101 gene expression may be associated with the expression changes of p21 and MMP2.
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ObjectiveTo investigate the role of Notch1 gene in xenografted human cutaneous squamous cell (SCL-1) carcinoma. MethodsFifteen nude mice were divided into three groups, including untreated group(inoculated with SCL-1 cells treated with phosphate buffered saline), empty vector group (inoculated with SCL-1 cells transfected with empty vector) and Notch1 group(inoculated with SCL-1 cells transfected with Notch1 expression vector). All the mice were inoculated with SCL-1 cells(1 x 108/ml) of0.2 ml. Then, the growth of xenografted tumor was observed every other day. Fifteen days later, the mice were sacrificed, tumor tissue was dissected and subjected to terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay for the detection of cell apoptosis, reverse-transcription(RT)-PCR and Western blot for the examination of mRNA and protein expressions of Notch1, bcl-2 and bax, respectively. ResultsThe proliferation of xenografted tumor in Notch1 group was obviously inhibited compared with the untreated group. The weight of xenografted tumor in Notch1 group was significantly lower than that in the untreated group and empty vector group (0.574 ± 0.219 g vs. 2.642 ± 0.404 g and 2.606 ± 0.512 g, F= 26.642, P＜ 0.01). TUNEL assay demonstrated that the number of apoptotic cells per 500 cells in tumor tissue specimens was(87 ± 9) in Notch1 group, evidently higher than that in the untreated group(8 ± 2) and empty vector group(10 ± 3) (F = 194.266, P ＜ 0.05 ). Further, RT-PCR and Western blot revealed that the mRNA and protein expressions of Notch1 and bax were significantly upregulated, but those of bcl-2 were markedly downregulated in the Notch 1 group, with significant difference among the three groups(all P ＜ 0.05). ConclusionsNotch 1 gene can inhibit the growth of xenogra ffted human cutaneous squamous cell(SCL-1) carcinoma and induce SCL-1 cell apoptosis likely by upregulating bax expression and downregulating bcl-2 expression.
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ObjectiveTo investigate the role and clinical pathological significance of PTTG and bFGF in cutaneous squamous cell carcinoma(CSCC).MethodsTissue specimens were collected from the lesions of 42 patients with CSCC and normal skin of 42 normal human controls.The protein and mRNA expressions of PTTG and bFGF were detected by immunohistochemistry and in situ hybridization in these specimens respectively.ResultsA significant increase was observed in the positive expression rates of PTTG and bFGF proteins[64.3％(27/42) vs.11.9％(5/42),73.8％(31/42) vs.21.4％(9/42),both P＜ 0.05] and mRNA [59.5％(25/42) vs.7.1％(3/42),75.0％(29/42) vs.16.7％(7/42),both P＜ 0.05] in the CSCC tissue specimens than in the control specimens.The protein and mRNA expressions of PTTG were positively correlated with those of bFGF(both P ＜ 0.05),and closely correlated with histological grade of CSCC (both P ＜0.05).ConclusionThe high expression of PTTG and bFGF may be associated with the initiation of CSCC.
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Objective To investigate the effect of pituitary tumor-transforming gene (PTTG)siRNA on the growth,invasion of,and expression of metastasis-related cytokines including matrix metalloproteinase-2 (MMP-2)and MMP-9 in xenografted human cutaneous squamous cell carcinoma in nude mice.Methods SCL-1 cells were subcutaneouslv inoculated into Balb/c nude mice to establish a xenograft model of human cutaneous squamous cell carcinoma.Then,15 mice bearing xenografted carcinoma were equally divided into 3 groups to be inoculated with phosphate buffer saline (PBS),control siRNA,and PTTG siRNA of 50 nmoI/L,respectively,ever),other day for 2 weeks.The size of xenograted carcinoma in these mice was measured every other day.At the end of 2-week treatment.the mice were killed followed by the evaluation of tumor weight,as well as the quantification of mRNA and protein expression of PTTG,MMP-2 and MMP-9 by reverse transcription (RT)-PCR and Western-blot,respectively.Results The xenograft model of human cutaneous squamous cell carcinoma was successfully established.The treatment with PTTG siRNA obviously inhibited the growth of the xenografted tumom and the expression of PTTG mRNA and protein compared with PBS and control siRNA (all P<0.05).In addition,the expression of MMP-2 and MMP-9 in xenografted tumors in PTTG siRNAtreated mice were significantly lower than those in PBS and control siRNA-treated mice.suggesting that PTTG siRNA evoked the decrease in invasive and metastatic ability of xenografted tumors.Conclusions PTTG siRNA can inhibit the growth of human cutaneous squamous cell carcinoma xenografts in nude mice,and downregulate the expression of invasion-and metastasis-related cytokines,including MMP-2 and MMP-9.
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Objective To study the effect of down-regulation of PTTG on the proliferation and migration of cutaneous squamous cell carcinoma cell line SCL-1 and its related mechanism. Methods SCL-1 cells were transfected with control siRNA or PTTG-targeting siRNA (PTTG-siRNA), or remained untransfected. After additional culture, the proliferation of SCL-1 cells as observed with cell counting kit-8 (CCK-8), and cell migration with Boyden chamber. Real-time PCR and Western blot were performed to detect the expression of matrix metalloproteinase 2 (MMP-2), MMP-9 and PTTG. Results The proliferation of SCL-1 cells transfected with PTTG-siRNA was markedly deccelarated in comparision with that of untransfected cells and those transfected with control siRNA (both P< 0.05). Real-time PCR and Western blot disclosed a significant decrease in the mRNA and protein expression of MMP-2, MMP-9 and PTTG in PTTG-siRNA-transfected SCL-1 cells compared with the other two groups of cells. As real-time PCR showed, the expressions of MMP-2, MMP-9 and PTTG in PTTG-siRNA-transfected SCL-1 cells were 0.8%, 23.2% and 21.3% of those in untransfected cells, respectively. Further more, the number of SCL-1 cells migrating through microporous membrane in the Boyden chamber was significantly smaller in PTTG-siRNA-transfected group than in untransfected group and control siRNA-trans-fected group (51.38 ± 4.71 vs 131.33 ± 6.12 and 127.72 ± 5.20, both P< 0.05). Conclusion The down-regulation of PTTG may deccelarate the proliferation and migration of SCL-1 cells and inhibit the expression of MMP-2 and MMP-9 in SCL-1 cells.
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Objective To investigate the effect of the exogenous fragile hisdidine triad(FHIT) gene on the proliferation and the apoptosis of cutaneous carcinoma cell line A431,and to explore the mechanism of tumor suppression by the FHIT gene.Methods The plasmids pcDNA3-FHIT and pcDNA3-vector were transfected into the cutaneous carcinoma cell line A431 without FHIT gene expression,and then the transfected cells were screened by G418 and the expression of FHIT was determined by the immunocytochemical staining technique.The effect of FHIT on the growth characteristics of cutaneous carcinoma cell line A431 was observed by MTT,colony forming test and flow cytometry.Results Stable FHIT gene expressing A431 cells were produced,the proliferation activity and colony forming capability of A431FHIT were suppressed,whereas the apoptosis was increased.All these differences between A431-FHIT cells and the two control groups of cutaneous carcinoma cells had statistical significance.Conclusion Transfecting the exogenous FHIT gene into cutaneous carcinoma cells line A431can suppress the proliferation of tumor cells,and can also induce apoptosis and cell cycle arrest.