Aim To investigate the inhibitory effects and mechanism of action of isoliensinine (IL) on the proliferation of porcine coronary arterial smooth muscle cells (CASMCs) induced by phenylephrine (Phen) and its mechanisms of action. Methods MTT assay, immunohistochemical method and Western blotting were adopted. Results IL (0.03-3 μmol·L-1) could inhibit the CASMCs proliferation induced by Phen (0.1 μmol·L-1) in a concentration-dependent manner. IL (0.1 μmol·L-1) antagonized Phen-induced overexpression of PDGF-β and bFGF from 0.545±0.026 and 0.47±0.03 to 0.458±0.019 and 0.376±0.017 (P<0.01, P<0.01). IL (0.1 μmol·L-1) also decreased c-fos, c-myc and hsp70 overexpression induced by Phen from 0.57±0.04, 0.44±0.04 and (173±36)% to 0.46±0.05, 0.372±0.021 and (115±35)% respectively (P<0.01, P<0.01, P<0.01). Conclusion IL exerted antiproliferative effect on CASMCs induced by phenylephrine, and its mechanisms were related to decrease the overexpression of growth factors (PDGF-β, bFGF), protooncogene (c-fos, c-myc) and hsp70.

Objective: To investigate the expression of EMS1 in laryngeal carcinoma and its clinical signifi-cance. Method:The expression of EMS1 protein was measured in 40 samples of, 40 samples of para carcinoma tis-sues (which were near to cutting margin of laryngeal carcinoma tissuse over 0. 5 cm) ,and 20 samples of normal la-ryngeal mucosa as controls by Flow Cytometere( Epics-XL Ⅱ ). Results:The quantity and percentage of EMS1 pro-tein expression in laryngeal carcinoma tissues was significantly higher than those in para carcinoma and in normal laryngeal mucosa tissues respectively(P<0. 05). There was no significant expression difference between the para carcinoma tissues and normal laryngeal mucosa tissues. There were positive correlation between the expressions of EMS1 protein and metastasis, pathological grade and clinical stage in laryngeal carcinoma. But there were not rela-tionship with patients' clinical classification, tumor size, smoking history, age and sex. Conclusion: The high ex-pression of EMS1 may contribute to the carcinogenesis and development of laryngeal carcinoma. The expression of EMS1 protein is an important index of judging differentiation, infiltration, metastasis and staging of laryngeal car-cinoma.

Objective To investigate the protective effects of ulinastatn on the lungs in patients with lung cancer undergoing lobectomy. Methods Forty ASA Ⅱ or Ⅲ patients with stage Ⅲ lung cancer, aged 50-64 yr weighing 53-70 kg undergoing lobectomy were randomly divided into 2 groups ( n = 20 each): control group (group C) and ulinastatin group (group U). In group U ulinastatin 10 000 U/kg in 20 ml normal saline was infused iv over 30 min immediately after induction of anesthesia. The patients were premedicated with diazepam 10 mg and scopolamine 0.3 mg im. Anesthesia was induced with midazolam 0.05 mg/kg, fentanyl 4 μg/kg, TCI of propofol (Cp 4 μg/ml) and vecuronium 0.12 mg/kg and maintained with TCI of propofol (Cp 2-3 μg/ml) and intermittent iv boluses of fentanyl and vecuronium. The patients were intubated with double-lumen tube. Correct position of the tube was checked with fiberoptic bronchoscope. One-lung ventilation (OLV) was performed (VT 6-8 ml/kg, RR 10-16 bpm, I:R 1:2, FiO2 100% ). PETCO2 was maintained at 35-45 mm Hg. Arterial blood samples were taken before anesthesia (T0, baseline), at 0.5 h and 1 h of OLV (T1, T2 ) and 4 h and 24 h after operation (T3, T4 )for blood gas analysis and determination of plasma TNF-α, IL-6 and IL-10 concentrations. Respiratory index (RI)was calculated. (RI= PA-a O2 /PaO2 ).Results Compared with the baseline values at To, plasma TNF-α and IL-6 concentrations and RI at T1-4 and plasma IL-10 concentrations at T1-3 were all significantly increased in group C,while in group U plasma TNF-α and IL-6 concentrations at T2,3 and plasma IL-10 concentrations and RI at T1-4 were all significantly increased ( P ＜ 0.05 ). Plasma TNF-α and IL-6 concentrations and RI were significantly lower while plasma IL-10 concentration was significantly higher in group U than in group C (P ＜ 0.05).Conclusion Ulinastatin 10 000 U/kg can effectively protect the lungs in patients with lung cancer undergoing lobectomy by attenuating systemic inflammatory response.

Aim To investigate the effects of rosmarinic acid on H2O2-induced apoptosis in rat VSMCs and its related mechanisms. Methods Flow cytometry was used to determine apoptotic rate of VSMCs. nuclear staining by acridine orange for morphologic change,MTT assay for cell viability and Western blot for expression of Bcl-2,Bax, fas and FasL. Results After being treated by H2O2(0,250,500,750 ?mol?L-1),apoptotic rate of VSMCs was 3.71%?0.56%,17.6%?6.92%,34.9%?2.55 %, and 85.6%?5.22% by FACS analysis,and VSMCs appeared nuclear condense and nuclear fragmentation after being treated by 500 ?mol?L-1 H2O2 for 24 hours,which were typical morphological changes of apoptosis. ① VSMCs treated with 500 ?mol?L-1 H2O2 for 24 hours had a significantly decrease on cell viability compared with control,and the apoptosis rate was increased to 35.7%?1.33%; Bcl-2 protein expression decreased,Bax protein expression increased, Bcl-2/Bax protein ratio decreased and Fas receptor and Fas ligand expression also increased.② Pre-incubation with rosmarinic acid(10, 20, 40 ?mol?L-1)for 30 min enhanced the cell viability, and decreased the apoptotic rate to 31.1%?1.38%,21.2%?1.18%,13.6%?0.51% in a dose-dependent manner. Moreover, Bcl-2/Bax protein ratio increased and Fas and FasL expression decreased. Conclusions ① Rosmarinic acid antagonizes H2O2-induced apoptosis of VSMCs. ② The antagonism of rosmarinic acid on apoptosis may be correlated with an increase Bcl-2/Bax protein ratio and a decrease expression of Fas and FasL protein.

objective To explore the correlation of single nucleotide polymorphism (SNP) frequency of adiponectin(APN)in patients with type Ⅱ diabetes(T2DM)and its complications,investigate whether the SNP is a risk factor of inheritance of T2DM,and to set up a highly efficient.accurate, economical and practical screening assay to detect the mutation of APN in clinical practice.Methods According to the diagnostic criteria of T2DM,patients with coronary heart disease(CHD),hypertension (HP),and diabetic nephropathy(NE)were recruited into this study.In simple,12DM group,T2DM-HP, T2DM-CHD.T2DM-NE and the control group.serum biochemistry items are measured.The technique of denaturing high-performance liquid chromatography(DHPLC)was used to detect SNPs of ANP gene.Results After all fragments amplified in the reglen of promoter of APN gene were compared with APN GeneID:9370 sequence recorded in GenBank,point mutation has been identified(-11377G/C).The frequency of genotypes of GG,GC,and CC are 5.16%,42.25%,52.58%and 3.4%.32.75%,63.85%,respectively in the groups of T2DM and control.The frequency of G allele was related to the incidence of T2DM,and is a risk factor of T2DM.The relative risk of GC to CC in developing T2DM is much high than that in the control group (OR=0.55).By comparing the clinical data of different groups of genotype in T2DM,it was observed that the genotype affected systolic blood pressure,BMI,abdominal circumference, and waist-buttock ratio(P=0.015).After optimizing the experimental conditions.it was found column temperature 6 0℃ was the best when using DHPLC technology to estimate SNP of APN gene.Conclusion SNP (-11377G/C) of APN gene G allele has a definite correlation with complications of hypertension in T2DM patients,and may contribute to the genetic risk for type 2 diabetes.

Objective To investigate the relationship between disease severity and enzyme-linked immunosorbent assay (ELISA) index values of anti-desmoglein (Dsg) 1 and-Dsg3 antibodies in 56 patients with pemphigus,and to characterize fluctuations in anti-Dsg1 and-Dsg3 antibodies in different forms of pemphigus.Methods Fifty-six patients with pemphigus (including 36 cases of pemphigus vulgaris (PV) and 20 cases of pemphigus foliaceus (PF)) were enrolled in this study.Blood samples were obtained from these patients before treatment and at the following four time points:when the condition was relieved and the taper of glucocorticoids began,the dose of glucocorticoids was tapered to half of their initial dose,the maintenance treatment started,and when the duration of maintenance treatment reached two years.ELISA was performed to determine the levels of anti-Dsg1 and-Dsg3 antibodies in these serum samples.Spearman correlation analysis was carried out to assess the relationship between disease severity and ELISA index values,and independent sample's t test to compare the levels of anti-Dsg antibodies among these time points.Results The severity of pemphigus was correlated with anti-Dsg antibody ELISA index values.Both anti-Dsg1 and anti-Dsg3 ELISA index values were significantly reduced at the remission of pemphigus compared with those before treatment (all P ＜ 0.01).At the end of the two-year maintenance treatment,10 (50％) patients with PF and 7 (19.4％) patients with PV became negative for anti-Dsg1 ELISA,whereas only 1 (2.7％) patient with PV became negative for anti-Dsg3 ELISA.Conclusions Anti-Dsg antibody ELISA index value is correlated with disease severity in patients with pemphigus,which may serve as a useful marker for assessing disease severity and activity as well as evaluating therapeutic efficacy.

OBJECTIVE: To investigate the expression of EMS1 in laryngeal carcinoma and its clinical significance. METHOD: The expression of EMS1 protein was measured in 40 samples of, 40 samples of para carcinoma tissues (which were near to cutting margin of laryngeal carcinoma tissue over 0.5 cm), and 20 samples of normal laryngeal mucosa as controls by Flow Cytometer (Epics-XL II). RESULT: The quantity and percentage of EMS1 protein expression in laryngeal carcinoma tissues was significantly higher than those in para carcinoma and in normal laryngeal mucosa tissues respectively (P<0.05). There was no significant expression difference between the para carcinoma tissues and normal laryngeal mucosa tissues. There were positive correlation between the expressions of EMS1 protein and metastasis, pathological grade and clinical stage in laryngeal carcinoma. But there were not relationship with patients' clinical classification, tumor size, smoking history, age and sex. CONCLUSION The high expression of EMS1 may contribute to the carcinogenesis and development of laryngeal carcinoma. The expression of EMS1 protein is an important index of judging differentiation, infiltration, metastasis and staging of laryngeal carcinoma.
Adult ; Aged ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cortactin ; metabolism ; Female ; Humans ; Laryngeal Mucosa ; metabolism ; pathology ; Laryngeal Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Staging

Objective To assess the antiangiogenic role of recombinant human endostatin combined with chemoradiotherapy and the capacity,and to explore the early tumor response as measured by comparing the change of MRI perfusion parameter.Methods From May 2012 to March 2013,22 locally advanced nasopharyngeal carcinoma patients who received recombinant human endostatin combined with chemoradiotherapy following induction chemotherapy,were included in the prospective study group.The other 25 patients,who received chemoradiotherapy following induction chemotherapy alone in the same period,were included in the control group.The perfusion parameters including blood volume(BV),blood flux(BF),mean transit time (MTT) were obtained by carrying out MR perfusion scanning at 3 time points:before induction chemotherapy,after induction chemotherapy,the end of concurrent chemoradiotherapy.Results Compared with before induction chemotherapy,the perfusion parameters including BV and BF obviously decreased in the study group (F =3.05,3.85,P ＜ 0.05).The parameter of MTT had no obviously change in the study group(P ＞0.05).In the control group,the change of BV,BF and MTT of nasopharyngeal lesions area during the treatment showed no significant difference (P ＞ 0.05).To make comparison between the two groups,at the end of concurrent chemoradiotherapy,BF of nasopharyngeal lesions area in the study group was 0.72 ± 0.56 and 1.92 ± 1.26 in the control group,the former showing significantly declined results (t =-3.056,P =0.012).Conclusions Recombinant human endostatin might be a good indicator of local tumor microvascular changes and the treatment-related toxicity could be tolerated.Magnetic resonance perfusion imaging maybe assessed the capacity of anti-angiogenesis therapy to induce early tumor response.Clinical trial registration Chinese clinical trial registry,ChiCRTONRC-12002394.

Objective To explore the regulatory role of hederagenin(HG)on glutamate(Glu)-in-duced ferroptosis and corresponding inflammatory responses in mouse hippocampal neuron HT22 cells and investigate its potential mechanisms.Methods HT22 cells were randomly divided into control,Glu and HG groups(n=3).The cells of the control group received no treatment,the cells of the Glu group were treated with 35 mmol/L Glu for 24 h to establish a cellular model of ferroptosis in Alzheimer's disease,and the cells of the HG group were treated with 0.5 μmol/L HG and 35 mmol/L Glu for 24 h simultaneously.FerroOrange fluorescent probe was used to de-tect intracellular Fe2+.The production of reactive oxygen species(ROS),mitochondrial membrane potential,and levels of inflammatory factors TNF-α,IL-1β and IL-6 in the cells were assessed.Finally,the expression of the key regulator of iron death,glutathione peroxidase 4(GPX4)was measured.Results Compared to the control group,the levels of intracellular Fe2+,ROS,TNF-α,IL-1β,and IL-6 were significantly elevated,while the mitochondrial membrane potential was obvi-ously reduced in the Glu group(P＜0.05,P＜0.01).The HG group had significantly decreased Fe2+,ROS,TNF-α,IL-1β,and IL-6 and enhanced mitochondrial membrane potential than the Glu group(P＜0.05,P＜0.01).The GPX4 expression was significantly lower in the Glu group than the control group(1.00±0.02 vs 0.46±0.04,P＜0.01),and was notably higher in the 0.5 and 1.0 μmol/L HG groups when compared to the Glu group(0.64±0.03 and 0.59±0.05 vs 0.46±0.04,P＜0.01).Conclusion HG inhibits ferroptosis by regulating GPX4 expression,and thereby effec-tively alleviates the inflammatory response.

Objective:To explore the effects of Onodera′s prognostic nutritional index (PNI) on the prognosis of locally advanced oropharyngeal squamous cell carcinoma (LA-OPSCC) after induction chemotherapy followed by sequential chemoradiotherapy.Methods:A retrospective analysis was conducted on the clinical data of 52 LA-OPSCC patients receiving induction chemotherapy followed by sequential chemoradiotherapy in The Affiliated Cancer Hospital of Guizhou Medical University during 2014-2018. The PNI values of all the patients at different treatment phases were statistically analyzed, and the ROC curve was employed to determine the optimal critical value of PNI. The patients in this study were divided into a well-nourished group ( n = 27) and a poorly-nourished group ( n = 25). The Kaplan-Meier method was used for survival analysis. The Cox proportional hazards model was utilized to analyze the relationships between different nutritional status and prognosis. Clinical features and adverse reactions were compared between the two groups. Results:The PNI values decreased significantly after radiotherapy, with an optimal critical value of 42.4. The 5-year overall survival (OS) and progression-free survival (PFS) of the well-nourished group (PNI ≥ 42.4) were 62.6% and 60.9%, respectively, which were significantly higher than those (30.1% and 29.7%) of the poorly-nourished group (PNI < 42.4, χ2 = 11.12, 5.74, P < 0.05). The multivariate analysis showed that PNI was an independent prognostic factor for the OS after radiotherapy ( HR = 2.752, 95% CI: 1.095-6.917, P = 0.031). The LA-OPSCC patients aged over 60 years or those who did not respond to induction chemotherapy accounted for a higher proportion of malnutrition after chemoradiotherapy ( χ2 = 4.89, 5.05, P < 0.05). Conclusions:PNI after radiotherapy can be used as a prognostic factor in the evaluation of LA-OPSCC patients receiving induction chemotherapy followed by sequential chemoradiotherapy. The LA-OPSCC patients aged over 60 years or those who do not respond to induction chemotherapy should receive more nutritional support during the chemoradiotherapy.