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Objective:To verify that hypoxia induces epithelial mesenchymal transition in HaCaT cells, and to observe the effect of LncRNA HOTAIR on epithelial mesenchymal transition in HaCaT cells.Methods:From Jan. 2018 to Dec. 2018 in Chinese Academy of Medical Science, HaCaT cells were divided into four groups: group A cultured under normoxia, group B cultured under hypoxia, group C transfected with sh-control cultured under hypoxia, and group D transfected with sh-LncRNA HOTAIR cultured under hypoxia. After 36 hours of culturing, the expression of E-cadherin and vimentin were detected using qPCR and Western blot. The migration of HaCaT cells was evaluated by Transwell assay.Results:The relative quantity of E-cadherin mRNA in the four groups were 3.076±0.271, 1.000±0.089, 1.024±0.222, and 2.595±0.085, while vimentin mRNAs were 1.002±0.183, 4.170±0.279, 4.111±0.477, and 2.412±0.134, respectively. In addition, the Transwll invasion assay showed that numbers of cell migration in the four groups were 32.70±3.93, 125.40±6.26, 120.10±6.79, and 58.24±7.06, respectively.Conclusions:The study suggests that hypoxia promotes epithelial mesenchymal transition in keratinocytes. Furthermore, downregulation of HOTAIR is noted to inhibit epithelial mesenchymal transition of keratinocytes under hypoxia condition.
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Objective:To observe the effect of LncRNA HOTAIR on the expression of HIF-1α in HaCat cell under low oxygen condition, and to explore the role of LncRNA HOTAIR in the pathogenesis and development of keloid.Methods:From Jan. 2018 to Dec. 2018 in Chinese Academy of Medical Science, recombinant plasmids were designed and constructed by specific shRNA-HOTAIR. After transfected HaCat cells with sh-LncRNA HOTAIR, RT-PCR was used to detect the expression level of HOTAIR. HaCat cells were cultured in different conditions and divided into four groups: group A cultured under normoxia condition, the other three groups cultured under hypoxia condition. Group C was transfected with sh-control before hypoxic culture, while group D were transfected with sh-LncRNA HOTAIR. After 24 hours′ culture, dual luciferase reporter gene assay was used to verify the relationship of LncRNA HOTAIR and HIF-1α. Expression of HIF-1α in four groups of HaCat cells were determined by Western blot analysis.Results:Recombinant plasmids of shRNA-HOTAIR were successfully constructed, and the HOTAIR expression was significantly decreased. The relative luciferase activity was 0.94±0.30 in group A, 20.39±1.15 in group B, 18.09±0.80 in group C and 3.04±1.15 in group D. The relative luciferase activity of group B was higher than those of group A ( t=29.03, P<0.05), and the difference was statistically significant ( P<0.05). According to the results of Western blotting, group B (1.19±0.07) and group D (1.15±0.06) had higher expression of HIF-1α than group A (0.56±0.29) and group C (0.37±0.38). Down-regulation of HOTAIR significantly inhibited the protein level of HIF-1α. Conclusions:LncRNA HOTAIR plays a positive role in upregulation of HIF-1α in HaCat cells under hypoxia condition. Thus LncRNA HOTAIR may take part in the pathogenesis and development of keloid through HIF-1α pathway.
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Objective To study the relationship between chemotherapeutic resistance to 5-fluorouracil (5-FU) and Golgi phosphoprotein 3 (GOLPH3) expression in human colon cancer cell line HT29.Methods HT29 cells were divided into four groups:control group,siRNA transfection group,experimental group 1 (30 μmol/L 5-FU),and experimental group 2 (siRNA-GOLPH3 + 30 μmol/L 5-FU).The silencing effect of GOLPH3 gene was detected by RT-PCR and Western blot.The tumor proliferation and formation ability of HT29 cell were measured respectively by MTT and plate colony formation assay.The protein expressions of GOLPH3,P-gp and β-catenin in HT29 cell were detected by Western blot.Results Compared with the control group,the expression level of GOLPH3 mRNA (1.000 ± 0.078 vs.0.147 ± 0.021,t =12.296,P ＜ 0.01) and protein(1.003 ± 0.235 vs.0.077 ± 0.399,t =6.723,P ＜ 0.01),the absorbance(A value) (1.000 ± 0.082 vs.0.769 ± 0.086,t =3.885,P ＜ 0.01) and the tumorigenesis (430 ±36 vs.297 ± 21,t =5.492,P ＜ 0.01) in transfection group significantly reduced.The A value (0.803 ± 0.101 vs.1.050 ± 0.140,t =2.855,P ＜ 0.05;0.242 ± 0.091 vs.1.050 ± 0.140,t =9.664,P ＜ 0.001) and the tumorigenesis (73 ± 8 vs.427 ± 29,t =20.363,P ＜ 0.001;305 ± 22 vs.427 ± 29,t =5.840,P ＜ 0.01) in experimental group 1 and experimental group 2 were significantly lower than that in the control group,while the A value (t =8.264,P ＜ 0.001) and the tumorigenesis (t =17.346,P ＜ 0.001) of experimental group 2 significantly decreased than that experimental group 1.Compared to the control group (0.967 ± 0.094,1.001 ± 0.199,1.000 ± 0.101),the expressions of GOLPH3,P-gp and β-catenin in experimental group 1 (3.634 ± 0.574,2.424 ± 0.261,2.324 ± 0.418) significantly upregulated (t =7.944,P ＜ 0.01;t =7.520,P ＜ 0.01;t =5.330,P ＜ 0.01),while the expressions of these proteins in experimental group 2(0.520 ±0.176,0.466 ±0.159,0.933 ±0.049) were significantly lower than that in experimental group 1 (t =8.983,P ＜ 0.01;t =11.106,P ＜ 0.001;t =5.724,P ＜ 0.01).Conclusion Expression of GOLPH3 is associated with resistance to 5-FU in colon cancer HT29 cell by activating Wnt/β-catenin signal pathway.
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Objective To investigate the clinical therapeutic effect of silver ions dressing combined with hy -drogel on diabetic foot .Methods 66 patients with diabetic foot admitted in our hospital during January 2012 and January 2014 were selected and randomly divided into control group and treatment group with 33 cases in each group . The control group was given conventional dressing therapy ,and the treatment group was given silver ions dressing com-bined with hydrogel treatment .The curative effects of both groups were then compared .Results The complete cure rate and cure rate of the treatment group were significantly higher than those in the control group , the time of debride-ment was longer than that in the control group , the growth of granulation tissue and epithelium time formation time was significantly shorter than that in the control group after treatment for 7d, the positive rate of bacteria was significantly lower than the control group .There were no obvious adverse reactions in the two groups during treatment .Conclusion In the treatment of diabetic foot , silver ions dressing combined with hydrogel treatment has better curative effect compared to traditional ones .
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The periods of the medical students' clinical practice is an important transition from medical theory to medical practice.According to the medical interns' expectations,such as the clinical tutor' level,hardware of the department,daily medical process and our department' s experiences,we grope for much better teaching methods that can meet medical interns' expectations and evaluate the new teaching methods.The basic clinical skills of medical students and the teaching level of gastrointestinal surgery department have been effectively improved.
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This study examined the effects of a recombinant adenovirus Ad-PTEN-EGFP on the proliferation of A549 cells, a human lung carcinoma cell line, in vitro and on the growth of the implanted tumors in the nude mice in vivo, explored the underlying mechanisms and evaluated the in vitro transfection efficiency of Ad-PTEN-EGFP into A549 cells. The expression of Ad-PTEN-EGFP in the A549 cells was determined. The proliferation and the apoptosis rates of the A549 cells with Ad-PTEN-EGFP transfection or not was detected by MTT and flow cytometry. Ad-PTEN-EGFP at different doses was injected intratumorally to the tumor-bearing mice induced by the A549 cells. Tumor sizes were measured on an alternate day. After all the mice were sacrificed, the implanted tumors were removed for routine histological examination, weight test, HE staining and immunohistochemical staining. The expressions of Bax, P16 and P53 in the tumor tissues and those of caspase-3, CD34 and VEGF in the mouse sera were detected. Tumor cell apoptosis was measured by TUNEL method. The results showed that the vitality of the A549 cells after transfection with Ad-PTEN-EGFP declined. The expression of green fluorescent protein was observed under fluorescent microscope. The transfection rate was in excess of 50%. The mRNA and protein expression of PTEN in the transfected cells was confirmed. The proliferation rate of the transfected cells was significantly decreased when compared with that of the non-transfected cells (P<0.05). The number of the apoptosis cells was increased in the transfected cells (P<0.05). The models of implanted tumors were successfully established by injection of the A549 cells in the flank of Balb/c nude mice. Administration of Ad-PTEN-EGFP to the tumor-bearing nude mice resulted in a suppression of tumor growth. There were statistically significant differences in the tumor weight and tumor volume between the Ad-PTEN-EGFP-treated group and the control groups (P<0.05). In contrast to those in the control groups, tumor tissues in the Ad-PTEN-EGFP-treated group were shown to have typical extensive vacuolar degeneration and massive hemorrhagic necrosis. Apoptotic bodies were also observed in the tumor cells. The expressions of Bax, caspase-3 and P16 were increased (P<0.05) while those of CD34, VEGF and P53 decreased (P<0.05) in the Ad-PTEN-EGFP-treated group. It is concluded that Ad-PTEN-EGFP could induce the apoptosis of the A549 cells and inhibit their proliferation. And it could also substantially suppress the tumor growth in the tumor-bearing nude mice and induce apoptosis of the tumor cells as well. These findings carry significant implications for adenovirus vector-based PTEN gene therapies for lung cancers.
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OBJECTIVE To explore how to realize the superior effect of Hospital Information System(HIS) in management of Nosocomial Infection(NI).METHODS Based on the platform of HIS,the prospective investigation,prevalence surveying,targeted surveillance and usage of antibiotics were performed.Drug-resistance was also monitored in real time.RESULTS Applying the HIS,infection control personnel could perform the overall surveillance on patients′ nosocomial infections.The hospital infection control process was optimized and time was saved for all medical personnel.CONCLUSIONS The application of the HIS enhances the hospital infection management level and decreases the incidence rate of NI,which may deserve to be generalized.
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The correlation between the expression of COX-2 and p53 protein in basal cell carcinoma (BCC) of eyelid and apoptosis was investigated. Specimens of BCC were collected from 40 cases (aged 28-68 y) at the Department of Pathology, Renmin Hospital of Wuhan University, and Department of Pathology, Zhongnan Hospital of Wuhan University during from 1999 to 2006. Five specimens of paracancerous tissues served as control group. Immunohistochemical staining was performed to detect the expression of COX-2 and p53 in the tissues. The average absorbance (A) and the average positive area rate of COX-2 and p53 protein were measured by image analysis. The positive area rate of COX-2 and p53 protein was analyzed by linear correlation analysis. It was found that COX-2 and p53 proteins were highly expressed in BCC of eyelid, and weakly expressed in paracancerous tissues. Image analysis revealed that the expression of COX-2 and p53 proteins in BCC of eyelid was significantly higher than that in paracancerous tissues (P<0.01). Spearman rank correlation analysis demonstrated a positive correlation between the expression of COX-2 and p53 (r=0.113, P=0.421). It was concluded that COX-2 can increase the expression of p53 protein, therefore suppressing apoptosis.
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Apoptosis , Carcinoma, Basal Cell/metabolism , Cyclooxygenase 2/metabolism , Eyelid Neoplasms/metabolism , Tumor Suppressor Protein p53/metabolismABSTRACT
Objective To observe the therapeutic effect of therapy of dispelling wind,cooling blood and tonifying kidney for refractory idiopathic thrombocytopenic purpura(RITP).Methods Fifteen RITP patients received therapy of dispelling wind,cooling blood and tonifying kidney.After treatment for 3 courses(18 weeks in total),the changes of platelet count and megakaryocyte maturation were observed.Results Of 15 RITP patiens,8 were clinically cured,4 markedly effective,2 effective,1 ineffective and the total effective rate was 93.3%.After treatment,the platelet count was increased(P
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[Objective] To explore the effect of wind-expelling and blood-cooling therapy in treating chronic idiopathic thrombocytopenia purpura (ITP) . [Methods] Thirteen cases of chronic ITP differentiated in TCM as heat-induced blood extravasation (group A) were treated with wind-expelling and blood-cooling therapy. The changes of peripheral T-lymphocyte subsets and natural killer (NK) cells before and after treatment were observed and compared with the normal volunteers (group B). [Results] Before treatment, peripheral T-lymphocyte subsets of CD3 and CD4 levels, CD4/CD8 ratio and NK cells count were lower in group A than those in group B while CD8 level was higher ( P
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[Objective] To observe the therapeutic effect of kidney-tonifying herbal medicine for chronic aplastic anemia (CAA) and its effect in regulating immune function. [Methods] Twenty-nine cases of CAA were randomized into 2 groups: group A (n = 15) was treated with routine western medicine plus Huosui Tablets and kidney-tonifying decoction for oral use, and Huosui Ointment for external application; group B (n = 14) was treated with routine western medicine only. Three months constituted one treatment course and the two groups were treated for over 2 courses. After treatment, the therapeutic effect was assessed and the changes of peripheral blood counting and immune function indexes were compared with those before treatment. [Results] After treatment, the total effective rate was 80.0% in group A and 42.9% in group B, the difference being significant (P 0.05) . [Conclusion] Kidney-tonifying herbal medicine is effective for the treatment of CAA and its therapeutic effect may be related to the regulation of immune function.
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AIM: The effects of low-glucose on long-term potentiation (LTP) in the CA1 region of hippocapal slices of immature (15-16 days old) and adult (56-63 days old) rats were examined. METHODS: The technique of electrophysiology was used, and the slopes of the field excitatory postsynaptic potentials (S-EPSP) were measured. RESULTS: When slices were exposed to glucose medium at concentrations of 3 or 1.5 mmol/L, S-EPSP decreased significantly. In the slices from adult rats, only short-term potentiation was elicited by high frequency stimulation in the medium of 3 or 1.5 mmol/L glucose. However, in the slices from immature rats, LTP was still induced in the medium of 3 mmol/L glucose. CONCLUSION: Low-glucose medium depressed the synaptic transmission. In terms of the synaptic plasticity, the low-glucose endurance in immature rats was stronger than that in adult rats.
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Objective To observe the influence of wind-dispelling therapy on therapeutic effect of blood-cooling and kidney-tonifying herbs for chronic idiopathic thrombocytopenic purpura(CITP),and to investigate its immunoregulatory effect.Methods Forty-two CITP patients were randomized into 2 groups:the treatment group(N=22) received therapy of dispelling wind,cooling blood and tonifying kidney,and the control group(N=20) received therapy of cooling blood and tonifying kidney.Six weeks constituted one treatment course,and the treatment lasted 3 courses.Serum interleukin 2(IL-2),soluble IL-2 receptor(sIL-2R),Fas protein,Fas-L protein,and platelet-associated immunoglobulin G(PAIgG) levels and lymphocyte subtypes were compared before and after treatment.Results In the treatment group,14 patients were clinically cured,5 markedly effective,2 effective,1 ineffective and the total effective rate was 95.5%;in the control group,8 patients were clinically cured,4 markedly effective,3 effective,5 ineffective and the total effective rate was 75.0%.The therapeutic effect in the treatment group was superior to that in the control group(P
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Objective To explore the therapeutic mechanism of blood- activating and blood- stasis- removing (BABAR) therapy for polycythemia vera (PV).Methods Ten first- visit or un- relieved outpatients and inpatients were treated with BABAR therapy for 12 weeks. Healthy volunteers served as the normal control. Before treatment and 1, 4, 8 and 12 weeks after treatment, the apoptotic rates of bone marrow mononuclear cells (BMMC) was analyzed with the TUNEL method. The expression of bcl- 2 and p53 was observed by the method of immunohistochemistry and their mRNA expression by the method of hybridization in site.Results One, four and eight weeks after treatment, the apoptotic rates of BMMC was increased as compared with those before treatment (P 0.05).The gene expression level of Bcl- 2 and p53 and their RNA expression level in the patients before treatment were higher than those in the normal control(P