Influencing factors of severe pneumonia in children with respiratory syncytial virus infection / 中国医师杂志

Objective:To explore the influencing factors of severe pneumonia in children with respiratory syncytial virus (RSV) infection.Methods:A retrospective case-control study was used to collect 210 children with RSV infected pneumonia admitted to Hebei Children′s Hospital from October 2017 to October 2020. Among them, 70 children with severe pneumonia were included in the severe pneumonia group, and 140 children with common pneumonia were included in the common pneumonia group; the baseline data and relevant laboratory indicators of the two groups were compared; Logistic regression was used to analyze the influencing factors of severe pneumonia in children infected with RSV.Results:The proportions of wheezing, congenital heart disease, respiratory failure, heart failure and pleural effusion of children in severe pneumonia group were higher than those in common pneumonia group, and the forced vital capacity (FVC) and forced expiratory volume in the first second (FEV 1) were lower than those in common pneumonia group (all P<0.05); the levels of C-reactive protein (CRP), CD8 + cells, RSV load and Beclin-1 in severe pneumonia group were higher than those in common pneumonia group, and the levels of CD4 + cells and 1, 25-dihydroxyvitamin D [1, 25-(OH) 2D] were lower than those in common pneumonia group (all P<0.05). After treatment, the levels of CRP, CD8 + cells and Beclin-1 in children with severe pneumonia were lower than those before treatment, and the levels of CD4 + cells and 1, 25-(OH) 2D were higher than those before treatment (all P<0.05). Multiple regression model analysis was established. The results showed that congenital heart disease, high CRP level, high CD8 + cells, high RSV load and high Beclin-1 level were risk factors for severe pneumonia in children with RSV infected pneumonia (all OR>1, P<0.05), and high CD4 + cells and 1, 25-(OH) 2D level were protective factors (all OR<1, P<0.05). Conclusions:Severe pneumonia in children with RSV infected pneumonia may be affected by congenital heart disease, CRP, CD4 + cells, CD8 + cells, 1, 25-(OH) 2D, RSV load and Beclin-1.

Characterization of the Eimeria tenella rhoptry protein with a nuclear localization sequence (EtROP30).

Rhoptry proteins (ROPs), secreted by specific rhoptry organelles of apicomplexan parasites, are determinants of parasite pathogenesis and sources of vaccine candidates. Twenty-eight ROPs of Eimeria tenella have been predicted by genomic approaches, and in the present study, E. tenella rhoptry protein 30 (EtROP30) was characterized. Subcellular localizations of EtROP30 in sporozoites and merozoites were in the apical complex and rhoptry-like bulb, suggesting that EtROP30 is a member of ROPs in E. tenella. Sequence analysis showed that EtROP30 contained an N-terminal secretory signal, a protein kinase domain with eight E. tenella-specific rhoptry kinase 1 subfamily (ROPK-Eten1) motifs, and a C-terminal nuclear localization sequence (NLS), making EtROP30 the only ROP that contains both a secretory signal and an NLS in E. tenella. Subsequent experiments showed that EtROP30 was a secreted protein in the sporozoite stage, relying on NLS for migration to the host nucleus. In addition, EtROP30 showed significantly higher expression levels in the parasite merozoite stage, indicating that EtROP30 plays a critical role during parasite reinvasion and development and may be a viable option as a vaccine candidate for anti-parasitic infection. The immunization protection efficacies of EtROP30 were evaluated. Significant improvements in mean body weight gain, reduction of cecum lesion score, and number of oocysts excreted were observed, indicating that EtROP30 has good immunogenicity against E. tenella. In the present study, a ROP of E. tenella with secretory and nuclear localization characteristics has been identified, and proved to be an effective vaccine candidate against this parasite.

Mechanism of tanshinone ⅡA inhibiting myocardial remodeling by Galectin-3 / 中华危重病急救医学

Objective:To explore the effect of tanshinone ⅡA on myocardial remodeling in ischemia/reperfusion (I/R)-induced heart failure of rodent model.Methods:① In vivo, 30 SD rats were randomly divided into sham operation, heart failure and tanshinone ⅡA treatment group, with 10 rats in each group. The I/R model was established by ligating the left coronary artery until ST segment elevation for 30 minutes, then the ligation was removed for 2 hours as reperfusion. In the sham operation group, the rat chest was opened without artery ligation. Three days after model establishment, tanshinone ⅡA (10 mg/kg) were given intraperitoneal injected in tanshinone ⅡA group for 9 weeks. In the other two groups, normal saline was administrated in the same way. The behavioral manifestations of the rats in each group were observed; hemodynamic indexes were evaluated; Masson staining was performed to observe the degree of myocardial fibrosis; enzyme linked immunosorbent assay (ELISA) was used to detect the content of Galectin-3 in myocardial tissue; quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was used to detect the expressions of collagenⅢ, collagenⅠ, matrix metalloproteinase 2 (MMP-2) and tissue inhibitor of metalloproteinase (TIMP-1). ② In vitro, rats primary cardiac fibroblasts were extracted and isolated, and divided into blank control group, angiotensinⅡ group (7-10 mmol/L angiotensinⅡ) and angiotensinⅡ+ tanshinoneⅡA group (7-10 mmol/L angiotensinⅡ+ 5-10 mmol/L tanshinone ⅡA). At 24 hours and 48 hours of culture, the cell proliferation in each group was detected by methyl thiazolyl tetrazolium (MTT); the expressions of collagenⅢ, collagenⅠ, MMP-2 and TIMP-1 were detected by qRT-PCR; the content of Galectin-3 in cardiac fibroblasts was detected by ELSIA. Results:① In vivo, the rats' activity status, hair conformity and food intake were ranked from good to bad in order of sham operation group, tanshinone ⅡA group and heart failure model group. Compared with the sham-operated group, the heart rate (HR) of the rats in the heart failure model group was significantly decreased and the heart function was significantly impaired. The mRNA and protein expression of collagenⅠ, collagenⅢ, TIMP-1 and Galectin-3 content were significantly increased, while the mRNA and protein expression of MMP-2 were significantly decreased. Compared with the heart failure model group, rats in the tanshinone ⅡA group showed significantly higher HR and improved cardiac function, significantly lower mRNA expression of collagenⅠ and collagenⅢ, significantly lower mRNA and protein expression of TIMP-1 and Galectin-3, and significantly higher mRNA and protein expression of MMP-2, and the most obvious changes were in the 9th weeks of modeling [collagenⅠ mRNA (2 -ΔΔCt): 4.70±1.19 vs. 10.21±1.62, collagenⅢ mRNA (2 -ΔΔCt): 3.03±0.46 vs. 13.84±1.93, TIMP-1 mRNA (2 -ΔΔCt): 1.90±0.19 vs. 4.55±0.43, TIMP-1/GAPDH: 0.33±0.04 vs. 0.67±0.05, Galectin-3 (ng/L): 489.93±79.30 vs. 821.72±94.09, MMP-2 mRNA (2 -ΔΔCt): 0.37±0.07 vs. 0.03±0.01, MMP-2/GAPDH: 0.69±0.09 vs. 0.21±0.04, all P < 0.05]. Masson staining showed that myocardial tissue fibrosis was obvious in the heart failure group, and the degree of fibrosis in the tanshinoneⅡA group was reduced. ② In vitro, compared with the blank control group, the proliferation rate, collagenⅠ, collagen Ⅲ and TIMP-1 expression and Galectin-3 content of myocardial fibroblasts were significantly increased, and MMP-2 expression was significantly decreased in the angiotensin group at 24 h and 48 h of culture. Compared with the angiotensin group, the proliferation rate of cardiac fibroblasts and the expression of collagenⅠ, collagen Ⅲ and TIMP-1 and the content of Galectin-3 were significantly decreased, and the expression of MMP-2 mRNA was significantly increased in the angiotensin + tanshinone ⅡA group, and the most significant changes were at 48 hours of culture [proliferation rate: (57.0±3.7)% vs. (67.0±2.4)%, collagenⅠmRNA (2 -ΔΔCt): 551.43±67.10 vs. 871.48±12.25, collagenⅢ mRNA (2 -ΔΔCt): 233.76±18.73 vs. 385.51±31.35, TIMP-1 mRNA (2 -ΔΔCt): 238.69±17.37 vs. 351.84±26.17, Galectin-3 (ng/L): 283.76±28.73 vs. 415.51±31.35, MMP-2 mRNA (2 -ΔΔCt): 108.54±12.10 vs. 51.47±6.25, all P < 0.05]. Conclusion:Tanshinone ⅡA can improve cardiac function, inhibit myocardial fibrosis and improve myocardial remodeling in rats with I/R-induced heart failure.

Meta-analysis of the effect of physical training on physiological function of adult renal transplant recipients / 中华肾脏病杂志

Objective To assess the impact of physical training on physiological function of adult renal transplant recipients by meta-analysis and to provide theoretical guidance for clinical practice.Methods Randomized controlled trials of physical training for the treatment of renal transplant recipients until October 2017 were searched in the database of Cochrane library,PubMed,Embase,Web of Science,Wanfang Data and CNKI.Data extracted from the literatures were analyzed with RevMan software (version 5.3).Results A total of 10 studies in 10 manuscripts met the inclusion criteria,and 557 cases were included.Meta-analysis results were as follows.Compared with the control group (routine drug therapy),the level of peak exercise oxygen uptake (peak VO2) was significantly increased in physical training group (routine drug therapy and physical training) (MD=2.40,95％ CI 0.15-4.64,P=0.04).However,there was no statistically significant difference in the change of blood lipid,blood pressure,hemoglobin and serum creatinine between the two groups (all P ＞0.05).Conclusions Physical training can improve cardio respiratory fitness of renal transplant recipients in the early stage,but it has no obvious effect on blood pressure,blood lipid,hemoglobin and blood creatinine.

High uric acid induces phenotypic transition of renal tubular cells via PI3K/Akt signaling pathway / 中华肾脏病杂志

Objective To investigate the effect and the mechanism of epithelial-mesenchymal transition (EMT) in renal tubular cells induced by uric acid.Methods Normal rat kidney tubular cell line (NRK-52E) were exposed to different concentrations of uric acid (100,200,400,600,800 μmol/L UA) for 48 hours to induce EMT.Morphological changes of the NRK-52E cells were examined under an inverted phase contrast microscope.The protein expression of E-cadherin,α-SMA,p-Akt and Akt were detected by Western blotting.The distribution of E-cadherin and α-SMA were detected by immunofluorescence.NRK-52E cells were pretreated by different concentrations of LY294002(0,2.5,5,10,15 μmol/L),the inhibitor of PI3K/p-Akt signaling pathway,and then processed by uric acid (400 μmol/L) for 48 hours.Western blotting was used to detect the protein expression of p-Akt and Akt.NRK-52E cells were then divided into four groups:normal group (N),uric acid group (UA),LY294002 group (LY),uric acid with LY294002 group (UA + LY).The protein expression of E-cadherin and α-SMA were detected by Western blotting,the distribution of E-cadherin,α-SMA and p-Akt were detected by immunofluorescence.Results There was abundant cellular expression of E-cadherin in unstimulated renal tubular cells whereas its expression was significantly decreased in uric acidstimulated cells (P ＜ 0.05).In addition,uric acid induced de novo expression of α-SMA in contrast to almost negative staining in untreated cells (P ＜ 0.05).p-Akt were obviously increased in high uric acid group (P ＜ 0.05) and Akt changed not significantly (P ＞ 0.05).NRK-52E cells transformed into elongated fibroblast-like cells from cuboidal clustered epithelial cells.These indicated that uric acid has induced EMT and activated PI3K/p-Akt signaling pathway in NRK-52E cells.However,the above effects of uric acid were abolished when p-Akt was blocked by the PI3K inhibitor (10,15 μmol/L LY294002),indicated that LY294002 has reversed the trend of EMT.Conclusions High uric acid induces phenotypic transition of renal tubular cells probably via activating PI3K/Akt signaling pathway.

Exploratory research of health services use of hierarchical medical system in elderly patients with type 2 diabetes mellitus in the community / 中华老年医学杂志

Objective To investigate the health services use of hierarchical medical system in elderly patients with type 2 diabetes mellitus in the community and to provide information for improving hierarchical medical system.Methods A survey of 684 T2DM patients aged 60 years and older was conducted in the Hua Mu Community Health Service Center in Shanghai from September 2014 to August 2016.And the data about health services visits for T2DM and its complications were collected in the latest one year.Based on different habits for patients receiving medical service,all patients were divided into two groups:the hierarchical treatment group and control group.All clinical and follow-up data were compared between two groups.Results 368 patients (53.8 ％)took on a priority in clinical visits to community health service center.There were no significant differences in age,gender,education,marriage,income,health insurance and duration of diabetes between the two groups(all P＞0.05).The acquired written health information about diet and exercise were much more in hierarchical treatment group than in the control group during one year.Although the rate of screening of diabetic nephropathy was similar between the two groups (P ＞ 0.05),the rate for retinopathy,neuropathy,vascular and diabetic foot was lower in hierarchical treatment group than in control group (all P ＜ 0.05).Moreover,the levels of glycosylated hemoglobin,urine albumin-to-creatinine ratio,total cholesterol,triglyceride,low-density lipoprotein cholesterol were similar between the two groups (all P＞0.05),but the level of high-density lipoprotein cholesterol in hierarchical treatment group was lower(P＜0.05).Furthermore,the rates of emergency visiting and hospitalization were lower in hierarchical treatment group than in the control group(both P＜0.001),and there was no significant difference in the average length of hospital stay between two groups (P ＞ 0.05).Conclusions Hierarchical medical system is favourable for the rational use of medical resources,but it is not yet quite perfect.Further improvements are still needed.

AFP-producing gastric cancer and hepatoid gastric cancer / 中华肿瘤杂志

AFP-producing gastric cancer(AFPGC) and hepatoid adenocarcinoma of the stomach (HAS) are two special subtypes of gastric cancer. There are both correlation and difference between them. AFPGC is usually identified as primary gastric cancer with serum AFP level more than 20 ng/ml or showed AFP positive staining by immunohistochemistry. The diagnosis of HAS is mainly dependent on the pathological character of hepatocellular carcinoma-like differentiation of gastric cancer. The morbidity of AFPGC and HAS are rather low, especially the incidence of HAS is about 1%. The prognoses of these two subtypes are poorer than that of common gastric adenocarcinoma, due to a high incidence rate of liver metastasis and lymph node metastasis. With the development of next-generation sequencing and other genomic technologies, gastric cancers, including these two rare subtypes, are now being investigated in more detail at the molecular level. Treatment remains the biggest challenge, early diagnosis and radical resection can dramatically improve patients′prognosis. Monitoring serum AFP and abdominal imaging examination during follow-up is important for early detection of liver metastasis. In combination with local treatment methods such as transarterial chemoembolization and radiofrequency ablation of liver may further extend patients′survival time. Targeted therapy owes a great potential value in the future.

The prognosis and clinicopathological characteristics of 70 gastric cancer patients with elevated serum AFP / 中华肿瘤杂志

Objective@#This study aimed to review the clinicopathological characteristics and prognosis of advanced gastric cancer patients with elevated serum Alpha-fetoprotein (AFP).@*Methods@#From August 2006 to May 2016, 70patients with advanced gastric cancer were enrolled retrospectively. All these patients had pathologically confirmed gastric adenocarcinoma, had a serum AFP level of more than 20 ng/ml, and received at least first-line treatment. The clinicopathological data and follow-up data were collected for analysis.@*Results@#Liver metastasis was more common in patients with AFP≥1 000 ng/ml, compared with AFP<1 000 ng/ml patients (78.6% vs 57.1%, P=0.064). In patients whose AFP level decreased ≥50% after first-line chemotherapy, the disease control rate and overall survival were both better than those in patients with AFP decreased <50%(96.3% vs 56.7%, P=0.001; 17.2 m vs 8.8 m P=0.007). The overall survival of all these 70 patients ranged from 0.5-50.0 months. 31 patients received chemotherapy only, and 39 patients received combined therapy modality. The overall survival of these two groups were similar(11.0 m and 15.6 m, respectively, P=0.070).@*Conclusions@#Serum AFP-elevated gastric cancer is a special subtype of gastric cancer. It′s a heterogeneous cancer with different clinical outcomes. The extent of decrease of serum AFP level during follow-up may be a sensitive prognostic and predictive biomarker. Liver metastasis were more common in these patients, and combined treatment may improve survival.

A case report of left coronary artery originated from the right coronary sinus in child / 天津医药

Anomalous coronary artery origin includes a variety of congenital coronary vascular variations. The left main coronary artery opening in the right coronary sinus is a rare and serious coronary artery malformation. The symptoms of angina, syncope and myocardial ischemia were found in this patient, but some patients showed no clinical symptoms and died suddenly after strenuous exercise. To improve the understanding of the disease early, the early surgery is the key to improve the prognosis.

Expressions of CD1a and CD68 in secondary keloid / 中华皮肤科杂志

ObjectiveTo analyze the distribution and density of Langerhans cells and dermal CD68 positive histiocytes in lesions of secondary keloid.MethodsTissue specimens were resected from the lesions of 30 patients with secondary keloid and normal skin of 14 human controls.Immunohistochemistry was performed to observe the expressions of CD68 and CD1a in these specimens.A micrometer was used to count the number of positively stained cells per unit area.The Student's t test was conducted for data analysis by using the SPSS software.ResultsThe density of CD1a+ Langerhans cells was (61 ± 49) cells/mm2 in the epidermis of secondary keloid lesions, (258 ± 61 ) cells/mm2 in the control epidermis,and(40 ± 65) cells/mm2 in the dermis of keloid lesions.CD68+ cells were absent in the epidermis of keloid lesions.Significant differences were observed in the density of CD1a+ Langerhans cells between the lesional and normal control epidermis(t =9.88,P ＜ 0.001 ) and in the percentage of CD68+ cells in nucleated cells between the superficial dermis of lesions and control skin(62％ ± 12％ vs.70％ ± 14％,t =2.66,P ＜ 0.05).The density of dermal CD68+ histiocytes was similar between the lesions and control skin ((287 ± 73) cells/mm2 vs.(290 ± 22) cell/mm2,t =0.02,P ＞ 0.05).Conclusions In keloid lesions,Langerhans cells decrease in the epidermis but increase in the dermis,CD68+ histiocytes are absent in the epidermis,and reduced in the dermis with a declined percentage in nucleated cells.

Effect of interferon-γand all-trans retinoic acid (ATRA) on the proliferation of and apoptosis in a human cutaneous squamous cell carcinoma cell line SCL-1 / 中华皮肤科杂志

Objective To investigate the in vitro effect of interferon-γ (IFN-γ) and ATRA on the morphological transition, proliferation of and apoptosis in a human cutaneous squamous cell carcinoma cell line SCL-1. Methods Cultured SCL-1 cells were divided into 6 groups to be treated with ATRA of 1 μmol/L, various concentrations ( 100, 500, 1000 U/ml) of IFN-γ, the combination of ATRA of 1 μmol/L and IFN-y of 1000 U/ml,respectively, or to remain untreated. MTT assay and flow cytometry were performed to evaluate the cell proliferation and apoptosis. The morphological features of apoptotic cells were observed by a transmission electron microscope (TEM) and inverted phase contrast microscope after 1% propidium iodide staining. Results IFN-γ could inhibit the proliferation of SCL-1 cells in a dose-dependent manner, and the most pronounced inhibitory effect was observed at a dose of 1000 U/ml . ATRA and IFN-γ induced an apoptosis in SCL-1 cells, and the early apoptosis rate was 4.84%, 11.96% and 18.71% in SCL-1 cells after treated with ATRA of 1 μmol/L, IFN-γ of 1000 U/ml and their combination, respectively. A series of morphological changes characteristic of apoptosis,such as bipolar changes, were observed in SCL-1 cells treated with ATRA and IFN-γ, with the presence of many early apoptotic cells, which showed a trend towards benign differentiation. Conclusions Within a certain concentration range, IFN-γcan promote the differentiation, but inhibit the proliferation of SCL-1 cells in a dose-dependent manner, and ATRA could enhance the effects of IFN-γ.

Effects of tumor necrosis factor (TNF)-α,interleukin(IL)-6 and interferon(IFN)-γ on the expression of CD68 in HaCaT cells / 中华皮肤科杂志

Objecfive To study the effects of some cytokines such as TNF-α,IL-6 and IFN-γ as well as lipopolysaccharide on CD68 expression in HaCaT cells.Methods Human HaCaT keratinocytes were randomly divided into natural proliferation group (without stimulation),IFN-γ-stimulated group,TNF-α-stimulated group,LPS-stimulated group and IL-6 stimulated group.The work concentration of TNF-α,IL-6,IFN-γ and LPS was 50 mg/L.HaCaT cells were collected after 24-hour treatment with the cytokines followed by the examination of CD68 expression with flow cytometry,immunohistochemistry and reverse transcription(RT)-PCR,respectively.Results Compared with untreated HaCaT cells,the count of CD68-positive cells was elevated in cells stimulated by TNF-α(t=3.60,P<0.01),IL-6(t=3.93,P<0.01),IFN-γ(t=2.38,P<0.05)and LPS(t=2.52,P<0.05),and the effect of TNF-α and IL-6 was stronger than that of IFN-γ and LPS.Among the four cytokines,only IL-6 enhanced the mean fluorescence intensity of CD68-positive cells (t=8.34,P<0.01).After 24-hour treatment with TNF-α,IFN-γ and IL-6,CD68 expression was observed in the cytoplasm and on the membrane of HaCaT cells and was stronger in cells treated with TNF-α and IL-6 than in those with the other cytokines.A significant increase was observed in the CD68 mRNA expression after 24-hour treatment with TNF-α (t=4.34,P<0.01),IL-6 (t=7.52,P<0.01)and IFN-γ (t=2.81,P<0.05);TNF-α and IL-6showed a stronger promotive effect than IFN-γ.Conclusion IL-6,TNF-α,IFN-γ and LPS can upregulate the CD68 expression in HaCaT cells.

Effect of local hyperthermia on Langerhans cells in the skin of a mouse model of contact hypersensitivity / 中华皮肤科杂志

Objective To investigate the effect of local hyperthermia on the morphology and quantity of Langerhans cells (LCs)at challenged skin sites of a mouse model of contact hypersensitivity.Methods Sixty mice were equally divided into 3 groups to be treated with local hyperthermia (37℃,39℃,41℃and 43℃)for 20 minutes at sensitization sites on the back of mice 3 days before (pre-heat group),concurrently with (concurrent-heat group)or 2 days after(post-heat group)sensitization respectively.Five mice which remained unsensitized and untreated served as the controls.Five days after the sensitization,the mice were challenged on the dorsal surface of right ears.Two days after the elicitation,the right ears were resected and immunohistochemistry was performed to observe the morphology and determine the quantity of LCs at challenged sites.Results With the rise in temperature,the number of LCs in the epidermis of ear skin decreased in pre-heat group(321.83±41.81,251.12±16.29,191.41±28.7,128.33±77.61 per square millimeter at 37 ℃,39 ℃,41℃and 43℃,respectively,P0.05)and in the post-heat group(320.83±113.6,398.33±31.91,437.83±29.78,477.25±86.79 per square millimeter at 37℃,39 ℃,41℃and 43℃,respectively,P<0.01).The dendrites of LCs increased in number and length when the temperature lose from 37 ℃ to 41℃,but slightly declined at 43℃.Conclusions Local hyperthermia at sensitization sites could affect the morphology and density of LCs at challenged sites,and the effect is likely associated with the severity of contact hypersensitivity.

Apoptosis in cutaneous squamous cell carcinoma cell line SCL-12 induced by baicalein and acitretin / 中华皮肤科杂志

Objective To investigate the effects of baicalein and acitretin on the apoptosis in a human cutaneous squamous cell carcinoma cell line, SCL-12. Methods Cultured SCL-12 cells were treated with different concentrations of baicalein (3.125, 6.25, 12.5 μmol/L) and acitretin (2.5, 5.0, 10.0 μ mol/L), alone or in combination, for 48 hours. Subsequently, cell proliferation was detected by MTT assay, and cell apoptosis by ELISA as well as annexin V-FITC and propidium iodide double staining. Real-time quantitative RT-PCR was used to detect the expression of Fas mRNA in SCL-12 cells. Results The cell proliferation of SCL-12 cells was inhibited by baicalein and acitretin alone or in combination. The combination of baicalein and acitretin at the three tested concentrations, except for that of baicalein at 3.125 μmol/L and acitretin at 2.5 μmol/L, more strongly inhibited the proliferation of SCL-12 cells compared with baicalein or acitretin alone, and the inhibitory effect was in a dose-dependent manner. The early apoptosis rate was 9.39% ± 1.52%, 20.86% ± 2.16%,36.85% ± 3.26% in SCL-12 cells treated with baicalein of 3.125 μmol/L, acitretin of 5.0 μmol/L alone and their combination, respectively, significantly higher than that in untreated cells (4.39% ± 0.64%, all P <0.05); the induction of apoptosis in SCL-12 cells by the combination of baicalein and acitretin was stronger than that by baicalein or acitretin alone (F = 138.44, P < 0.05). Baicalein and acitretin alone or in combination significantly increased the mRNA expression of Fas in SCL-12 cells, and the effect of their combination was stronger than that of baicalein or acitretin alone. Conclusions Baicalein and aeitretin could inhibit the growth of and induce the apoptosis in SCL-12 cells, and the effect is enhanced by the combination of baicalein and acitretin, which may be associated with the upregulation of Fas expression in SCL-12 cells.

Expression of survivin and bcl-2 in human skin squamous cell carcinoma lesions and cell line SCL-1 / 中华皮肤科杂志

Objective To investigate the expression of survivin and bcl-2 in human squamous cell carcinoma (SCC) lesions and cell line SCL-1. Methods Tissue samples from 60 patients with SCC and 10 normal human controls were immunohistochemically stained to detect the expressions of survivin and bcl-2.Western blot was used to measure the expressions of bcl-2 and survivin proteins in HaCaT human keratinocytes and SCL-1 human squamous cell carcinoma cells. Results In normal control tissues, there was no expressions of survivin or bcl-2, while in SCC, the expression rates of bcl-2 and survivin were 70% and 60%, respectively,and there was no statistical correlation between the expressions of bcl-2 and survivin (P >0.05). Neither the expression of survivin nor that of bcl-2 was correlated to patients' age, gender or lesional site (all P >0.05). A statistical correlation was observed between the pathological stage in patients and expression of bcl-2 as well as between lymph node metastasis and expression of survivin (both P < 0.05). Western blot analysis revealed a significant increase in the expression of survivin and bcl-2 in SCL-1 cells compared with HaCaT cells. Con-clusion In SCC, survivin and bcl-2 seem to play their roles via different anti-apoptotic pathways.

Development and primary application of a real-time quantitative polymerase chain reaction for the detection of human breast cancer related novel gene-Metadherin expression / 中华内分泌外科杂志

Objective To establish a quantitative SYBR GreenⅠreal-time PCR method for detection of Metadherin (MTDH) gene expression level in breast cancer, and to investigate the relationship between MTDH mRNA expression level and the clinicopathological characters. Methods Real-time PCR was employed to determine the expression level of MTDH mRNA in peripheral blood of 80 specimens of breast cancer patients and normal females. Results In the 80 specimens, MTDH mRNA was positively expressed in the peripheral blood of 61 cases of breast cancer patients while negatively expressed in the 19 cases of normal females. Among the 61 breast cancer patients, MTDH mRNA showed high expression in 34 cases, accounting for 55.7%, and showed low expression in 27 cases, accounting for 44.3%. Both of the differences in expression rate has statistic significance (Ratio = 2.02 ± 0.81, P<0.05). MTDH expression relative to GAPDH expression in the peripheral blood of breast cancer patients was 1.15 ± 0.36. MTDH mRNA expression has no connection with age, estrogen and progesterone receptors , as well as HER-2.(P＞0.05). There is statistical difference for MTDH mRNA expression level between the group with lymph node metastasis and the group without lymph node metastasis (Ratio=2.02 ± 0.81,P<0.05). MTDH mRNA expression level changed between clinicopathological stage I, II and III, IV. Conclusion The established SYBR Green Ⅰ quantitative real-time PCR method can successfully detect the expression level of MTDH mRNA, which may be closely related to the occurrence and development of breast cancer.

Cell activity of human de-epidermized dermis and its Characteristics of tissue Structure / 中国组织工程研究

BACKGROUND:Study confirmed that the de-epidermized dermis(DED)can be used as dermal substitute and may form epidermal structure after incubating keratinocytes.However,the cell biological activity,tissue structure characteristics and the basement membrane component analysis of dermal substitute have been reported less.OBJECTIVE:To investigate the cell activity and the tissue structure characteristics of DED.METHODS:Skin flap was treated with 56℃ phosphate buffered solution to remove the epidermis,and the dermal cell components were deleted by freezing and thawing with liquid nitrogen to obtain DED.The DED cell activity was detected with tissue culture method,hematoxylin nuclear staining was used to determine the DED cell nuclei,and vimentin immunohistochemistry was applied for fibroblast determinations.The basement membrane and its components were detected using Periodic Acid-Schiff staining and collagen type Ⅳ immunohistochemistry.Van Gieson stain,Weigart stain and those double staining were respectively used to determine DED collagen fibers and elastic fibers.The DED ultrastructure was observed under transmission and scanning electron microscope.RESULTS AND CONCLUSlON:Using tissue culture method,the cultured DED did not exhibit cell growth at 2 weeks.Hematoxylin-eosin staining showed no nuclear in DED,vimentin immunohistochemistry showed no vimentin expressed in DED.Van Gieson staining showed DED collagen fibers were stained as rose red,Weigert staining showed DED elastic fibers were stained as pu rplish black double staining further demonstrated uniform arrangement of collagen fibers and elastic fibers.DED surface and the remaining appendages were strongly positive for Periodic Acid-Schiff staining,and type Ⅳ collagen expression was significant.Transmission and scanning electron microscope results showed that,the DED elastic fibers and collagen overlap arranged with pore intervals,they intercrossed into a network.There is no living cell component in DED,dermal matrix surface and appending organ luminal wall still retain glycogen,type Ⅳ collagen and other basement membrane components,dermal matrix is rich in collagen and elastic fibers.it is a three-dimensional collagen matrix similar to in vivo dermis.

Effects of hyperthermia on the expression of E6 and E7 genes of human papillomavirus type 6 and 11 / 中华皮肤科杂志

Objective To investigate the effect of hyperthermia on the expression of E6 and E7 genes of human papillomavirus (HPV) type 6 and 11 in HPV-infected human skin. Methods Tissue samples were obtained from the lesions of condyloma accuminatum (CA) in 6 patients after informed consent. Each sample was divided into 4 parts: one was embedded and directly stored at -80 ℃; the other 3 parts were placed in culture medium and the surface of the samples was irradiated for 30 minutes with a thermotherapy apparatus at 37℃, 42 ℃, 45 ℃, respectively, then the samples were taken out and stored at -80 ℃. RNA was extracted from the specimens, real time quantitative PCR (qPCR) was performed to detect the expression of E6 and E7 genes of HPV-6 and -11. Results Of the 6 patients, 2 were infected with HPV-6 and -11 respectively, 4 with both HPV-6 and HPV-11. The expression of E6 and E7 mRNA decreased with the increase in irradiation temperature. The relative mRNA expression levels at 37 ℃, 42 ℃ and 45 ℃ were 1.00 ± 0.00, 0.61 ± 0.17, 0.27 ± 0.15, respectively, for HPV-6 E6 gene, 1.00 ± 0.00, 0.56 ± 0.21, 0.16 ± 0.11 respectively, for HPV-6 E7 gene, 1.00 ± 0.00, 0.60 ± 0.22, 0.16 ± 0.08, respectively, for HPV-I1 E6 gene, 1.00 ± 0.00, 0.55 ± 0.15, 0.24 ± 0.06, respectively, for HPV-11 E7 gene; statistical difference was noted among them between the specimens irradiated at different temperature (all P < 0.01). Conclusion Hyperthermia can remarkably suppress the expression of HPV-6/I 1 E6 and E7 genes, which may be a possible mechanism under the regression of warts induced by local hyperthermia.

Effects of local hyperthermia on phosphatidylinositol 3-kinase gene expression in Langerhans cells from human papillomavirus(HPV)-infected skin / 中华皮肤科杂志

Objective To investigate the effect of local hyperthermia on the mRNA expression of phosphatidylinositol 3-kinase(P13-K)in Langerhans cells(LCs)transmigrating from HPV-infected skin.Methods Tissue samples were collected from 5 female patients with condyloma acuminatum(CA)and 5 normal human controls.then equally divided into three parts to receive local hyoerthermia treatment at 37℃,42 ℃ and 45℃,respectively,for 30 minutes.After another 12-hour incubation in RPMI 1640 culture medium,transmigrating cells were collected and LCs were purified.Real-time quantitative reverse transcription-PCR was performed to detect the mRNA expressions of P13-K p85a and p110α in purified CD1a~+ LCs from these tissue samples.Results In CD1a~+ LCs transmigrating from normal skin and lesions of CA.the expressions of P13-K p85a and P110α mRNA decreased with the increase of temperature.Atier local hyperthermia treatment of normal skin at 37℃,42℃,45℃,the relative mRNA expression level was 1.00±0.00,0.78±0.13,0.54±0.17,respectively.for P13-K p85α in transmigrating LCs,1.00±0.00,0.80±0.11,0.61±0.12,respectively.for P13-K P110α;there was a significant difierence among the three temperatures in bOth parameters(both P＜0.01).In the case of lesions of CA.the relative mRNA expression level of P13-K p85αand p110α was 1.00±0.00 and 1.00±0.00 under treatment at 37℃,0.20±0.11 and 0.49±0.21 at 42℃.0.1±0.08 and 0.09±0.03 at 45℃.respectively;significant difierence was also noted among the three treatment temperatures(both P＜0.01).The decrease in P13-K P110α mRNA expressions under hyperthermia treatment at 42 ℃ and 45℃ compared with those at 37℃was greater in LCs from CA lesions than that from normal skin.Conclusions Local hyoerthermia could remarkably inhibit the expression of P13-K genes.which may enhance immunity and favor elimination of HPV.