OBJECTIVETo investigate the effects of oral cancer-associated fibroblasts (CAFs) on lymphangiogenesis in oral squamous cell carcinoma (OSCC).

METHODSCAFs and normal fibroblasts (NFs) were obtained from the tissues of patients with OSCC who did not receive radio-chemotherapy before operation. And the CAFs and NFs were isolated by method of tissue block and identified by immunohistochemical staining. The effects of CAFs (group A) and NFs (group B) to human lymphatic endothelial cells (HLEC) were detected by using a 24-multiwell transwell cell culture chamber. DMEM sugar medium was as blank control group. The number of proliferative, migratory, invasive and tubes of HLEC were counted under inverted phase contrast microscope.

RESULTSThe proliferative number of HLEC of group A for 96, 144, 196 h was significantly higher than that of group B and blank control group, group B higher than blank control group (P<0.01). The migratory and invasive number of HLEC of group A for 96 h was significantly higher than that of group B and blank control group, group B higher than blank control group (P<0.01). The number of tube formation of HLEC of group A for 24 h was significantly higher than that of group B and blank control group, group B higher than blank control group (P<0.01).

CONCLUSIONCAFs promote HLEC's proliferation, migration, invasion, tube formation, and these effects are stronger than NFs.

Carcinoma, Squamous Cell ; Cell Culture Techniques ; Cell Movement ; Cell Proliferation ; Endothelial Cells ; Fibroblasts ; physiology ; Humans ; Mouth Neoplasms ; pathology

Objective To study the expression of interleukin (IL-17A) in primary liver cancer (PHC) tissue and its clinical significance.Methods Reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry was performed respectively to detect the expression of IL-17A mRNA and CD34 in tumor tissues from 37 patients with PHC.Murine H22 cells cultured in vitro were exposed to IL-17A at different dosages (0.1,0.5,1.0,5.0,10,50,100,500,1000 ng/ml) and the cell proliferation was assayed by MTT.IL-17A was administered to the mice transplanted with H22 cancer cells via caudal vein and the tumor volume was measured by vernier caliper.Immunohistochemistry was used to detect the CD31 expression in H22 cancer tissues.Results IL-17A mRNA was detected in 26 of the 37 samples of primary liver cancer.The microvessel densities in IL-17A-positive samples and IL-17A-negative samples were 66.6 ± 2.5 and 26.7--2.5,respectively.The difference between two groups was significant (P＜0.01).The proliferation of H22 cells exposed to various dosages of IL-17A was not different (P＞0.05).The volume of murine tumor tissue in animals treated with IL-17A was (843.6± 90.9) mm3 and in untreated animals was (198.7±24.4) mm3 (P＜0.01).The microvessel density in treated group and control group was 71.9± 6.8 and 33.3 ± 2.9,respectively (P＜0.01).Conclusions The expression of IL-17A could be detected in a considerable proportion of primary liver cancers and correlated with angiogenesis.Exogenous IL-17A could not accelerate H22 cell proliferation in vitro but in vivo probably via enhancing angiogenesis.

Objective To explore significance of the T lymphacyte subgroup in cases with multiple sclerosis(MS).Methods Indirect immunofluorescence assay and flow cytometry were used to evaluate the presence of CD_4~+,CD_8~+ and T lymphocyte in the sera and CSF in patients with MS treated with glucocorticoid in the active stage.Results The CD_4~+,CD_8~+ and T lymphocyte in the peripheral blood of the patients in comparison with those of the control group decreased whereas their CD_4~+/CD_8~+ increased(P

Objective To explore the effectiveness of long-acting antibacterial material that was used as an intervention in oral care cavity nursing in patients with endotracheal intubation and mechanical ventilation. Methods Patients admitted to Xuzhou Medical College Hospital neurosurgery intensive care unit between 1st December 2013 and 31st August 2014 requiring intubation and mechanical ventilation were enrolled into the study. There were 100 cases randomly selected and were assigned into control group and experimental group by random number table. Each contained 50 cases. Control group used saline for oral care by brushing and irrigation;experimental group used antimicrobial material oral spray in oral care on the top of the method used in control group. The mouth odor, mouth infection rate and ventilator-associated pneumonia (VAP) rate were used to determine. Results About oral odor, mouth infection rate, and VAP rate, the difference of incidences between experimental group and the control group was significant (χ2=4.504-13.728, P <0.05). Conclusions Antimicrobial material can effectively improve oral health in intubated and mechanical ventilated patients. It slows oral acidic environment change, reduces oral infections early onset VAP incidence. And it is worth promoting its use.

AIM: To investigate the effects of PAR-2 agonist peptide on the proliferation and cytosolic calcium concentration ([Ca~(2+)]_c) in human hepatoma cells HepG2. METHODS: Human hepatoma cell line HepG2 was cultured. The cells were treated with PAR-2 agonist peptide SLIGKV-NH_2 and the reverse PAR-2 agonist peptide VKGILS-NH_2, respectively. The [Ca~(2+)]_c of hepatoma cells were measured by microfluorimetric techniques based on calcium indicator fura-2/AM. The influences on proliferation of hepatoma cells were determined by MTT method. The changes of cell cycle were evaluated by flow cytometry, and the changes of cyclin D1 mRNA expression were detected by RT-PCR. RESULTS: After treated with 50 μmol/L SLIGKV-NH_2, a rapid rise of [Ca~(2+)]_c in HepG2 cells was induced (P<0.01), percent S phase, G_2/M phase and proliferation index (PI) of HepG2 cells were elevated (P<0.01), and cyclin D1 mRNA expression was significantly upregulated (P<0.01). The proliferation rates of HepG2 cells treated with 1-50 μmol/L SLIGKV-NH_2 were significantly increased, and the effect was in a dose-dependent manner (P<0.01 or P<0.05). No statistical significance of the difference between VKGILS-NH_2 and control group was observed (P>0.05). CONCLUSION: PAR-2 agonist peptide induces the rise of [Ca~(2+)]_c in HepG2 cells, upregulates the expression of cyclin D1 mRNA, accelerates the progress of cell cycle, promotes the synthesis of DNA and the proliferation of hepatoma cells via activating PAR-2 in vitro.

Objective To evaluate the influence of ultrasound-mediated contrast agent microbubbles carrying kallidinogenase targeted therapy on neurogenesis and angiogenesis after experimental acute cerebral infarction.Methods Kallidinogenase-loaded microbubbles were prepared using mechanical shaking method.Middle cerebral artery occlusion (MCAO) model was established in male Wistar rats by sutureoccluded method.MCAO rats (n =120) were randomly divided into 5 groups:ultrasound-mediated kallidinogenase-loaded microbubbles group (group 1),ultrasound-mediated microbubbles group (group 2),ultrasound-mediated saline group (group 3),kallidinogenase group (group 4),saline group (group 5).Medication was given through tail vein daily for 2-6 consecutive days starting 24 h after MCAO.Ultrasound groups (groups 1,2 and 3) were given 2 MHz pulse ultrasonic irradiation on the ischemia lateral skull for 10 min after injection.Cell proliferation was examined using 5'-bromo-2'-deoxyuridine (BrdU,50 mg/kg).Infarction volume and neurological function were evaluated on the 3rd,7th day after MCAO respectively.Doublecortin (DCX) + cells in the subventricular zone and laminin + in the peri-infarction region were observed at the same time.Results The number of DCX + cells in the subventricular zone (under 20 × ocular) of groups 1-5 was 251.8 ± 13.1,125.7 ± 11.6,130.2 ± 13.7,234.5 ± 12.4 and 123.7 ± 10.0 respectively.The percentage of laminin + cells in the peri-infarction region (× 10 objective) of groups 1-5 was 10.0％ ± 0.8％,5.2％ ± 0.7％,5.0％ ± 1.0％,8.0％ ± 1.8％ and 5.0％ ± 0.9％ respectively.The number of DCX + cells in the subventricular zone and laminin + cells in the peri-infarction region of the group 1 and the group 4 was significantly more on the 7th day after MCAO compared with those of the other three groups (DCX:t values of group 1 vs groups 2,3,5 were 17.88,17.17 and 18.16,all P ＜ 0.01 ; t values of group 4 vs groups 2,3,5 were 15.42,14.78 and 15.70,all P ＜0.01.Laminin:t values of group 1 vs groups 2,3,5 were 7.01,6.71 and 7.11,all P ＜ 0.01 ; t values of group 4 vs groups 2,3,5 were 4.23,3.94 and 4.33,all P ＜0.01).Moreover,the number of DCX+ cells in the subventricular zone (t =2.46,P ＜ 0.05) and laminin + cells in the peri-infarction region (t =2.78,P ＜ 0.05) of the group 1 was much more than those of the group 4.Neurologic scores on the 7th day of groups 1-5 were 1.00 (0.75,1.25),2.0 (2.00,3.00),2.0 (1.00,2.00),1.5 (0.75,2.00),2.0 (2.00,2.50).Compared with other four groups,group 1 showed better functional improvement after stroke (U values of group 1 vs groups 2-5 were 2.0,4.0,7.5 and 2.5,all P ＜ 0.05).While there was no significant difference in infarction volume among five groups at all the time points after MCAO.Conclusions Our study demonstrates ultrasound-mediated kallidinogenase-loaded contrast agent microbubbles targeted therapy promotes neuroblasts proliferation and vascular regeneration compared with mediated and non-medicine-loaded microbubbles therapy,which attributes to functional improvement after MCAO.Therefore,ultrasound-mediated ultrasound contrast agent microbubbles carrying drugs targeted therapy may have a perspective on ischemic stroke.

Objective To evaluate the predicting value of the marker of endothelial injury: plasma matrix metalloproteinase-9( MMP-9) and high sensitivity C-reactive protein( hsCRP) level on the progression of acute anterior circulating territory infarction progressing to malignant middle cerebral artery infarction (m-MCAI). Methods 90 patients with acute anterior circulating territory infarction, in which 46 patients progressed to m-MCAI, were collected and sampled consecutively. The plasma MMP-9 and hsCRP of all patients were determined by ELISA and immunotur-bidimetry,respectively,at admission. And the clinical characters and cranial CT features of the patients were analyzed. Results At admission,the plasma MMP-9 level in the patients with m-MCAI(242.0 ±58.0)ng/ml was significantly higher than that in the patients with non m-MCAI( 169.0 ± 50.0) ng/ml( P < 0. 01) ,the plasma hsCRP level in the patients with m-MCAI(6.25 ±1.2) ng/ml was significantly higher than that in the patients with non m-MCAI( 1.55 ± 0.9) mg/ml( P <0. 01).Conclusion The increased level of plasma MMP-9 and hsCRP could be predictors for the m-MCAI proceeding.

This study is to investigate the mechanism of human promyelocytic leukemia HL-60 cells proliferation induced by alteronol in vitro. Human promyelocytic leukemia HL-60 cells cultured in vitro were treated with different concentrations of alteronol. Inhibition rate was detected by SRB assay. Cellular morphological changes were observed by Hoechst and AO/EB (acridine orange/ethidium bromide dye) staining. The apoptosis rate was determined by Annexin V-FITC/PI assay. Cell cycle distribution was determined by flow cytometry. Western blotting analysis was carried out to determine the cell cycle related proteins. The proliferation of HL-60 cells treated with alteronol was inhibited in a concentration-dependent manner. Based on cell viability assay, observation on cell morphology and apoptosis rate, it confirmed that alteronol played an obvious role in proliferation inhibition of human promyelocytic leukemia HL-60 cells, but it did not induce apoptosis in human promyelocytic leukemia HL-60 cells in different concentrations groups. Alteronol could effectively inhibit the proliferation of human promyelocytic leukemia HL-60 cells inducing cell cycle arrest at G1 phase, as well as, alteration expression of cell cycle proteins level of CyclinD1 and pRb.

Objective To evaluate the cardioprotection induced by ischemic preconditioning ( IPC) and hypoxic preconditioning ( HPC) which were carried out by using the double cardiopulmonary bypass ( CPB) circuits in in vivo hearts of dogs. Methods Eighteen healthy male dogs, weighing 17?5-24?5 kg, aged 13-24 months, were divided into 3 groups ( n=6 each) using a random number table: my?ocardial ischemia?reperfusion group (group I∕R), IPC group and HPC group. The double CPB circuits were established as follows: systemic and coronary circulation, and independent systemic and coronary cir?culation was carried out. In group IPC, the aorta was clamped, and the coronary circulation pump was sus?pended for 5 min followed by 5 min opening, repeating for 3 cycles. In group HPC, the aorta was clamped, the coronary circulation was started, and pure nitrogen was insufflated for 5 min followed by 5 min of oxygen insufflation, repeating for 3 cycles. CPB was performed for 1 h starting from the time point immediately after IPC or HPC. Before splitting of sternum ( T1 ) , after establishment of double CPB circuits ( T2 ) , at the end of preconditioning ( T3 ) , and at 60 and 120 min after restoration of spontaneous heart
beat ( T4,5 ) , heat rate, mean arterial pressure, central venous pressure, left ventricular end?systolic pres?sure, left ventricular end?diastolic pressure and the maximum rate of increase∕decrease of left ventricular pressure were recorded. Blood samples were collected from the right internal jugular vein at T1 and T4,5 for determination of serum cardiac troponin I concentrations. The animals were sacrificed after determination of the parameter or after blood sampling at T5 , myocardial specimens were obtained for examination of the ul?trastructure and for detection of apoptosis in cardiomyocytes, and apoptosis index was calculated. Before aortic clamping, immediately after aortic unclamping and at 30 min after aortic unclamping, myocardial specimens were obtained for determination of ATP contents in cardiomyocytes. Results Compared with I∕R group, left ventricular end?systolic pressure was significantly increased, and the serum cardiac troponin I concentrations were significantly decreased at T4,5 , the myocardial ATP contents were significantly in?creased immediately after aortic unclamping and at 30 min after aortic unclamping, apoptosis index was sig?nificantly decreased ( P<0?05 or 0?01) , and the pathological changes were significantly attenuated in IPC and HPC groups. Compared with group IPC, the myocardial ATP contents were significantly increased (P<0?05), and the pathological changes were attenuated in group HPC. Conclusion Both HPC and IPC can exert cardioprotection when carried out by using the double CPB circuits, and HPC provides better cardioprotection than IPC in in vivo hearts of dogs.

Objective:To investigate the MRI features of the primary sinonasal malignant melanoma (SMM) and evaluate the signal pattern based on T 1WI and T 2WI, in order to improve the diagnostic accuracy of SMM. Methods:The MRI findings of 63 SMM cases confirmed by pathology from April 2007 to November 2018 at Beijing Tongren Hospital, Capital Medical University were analyzed retrospectively. The signal intensity of malignant melanoma was classified into four types(Ⅰ—Ⅳ) according to the proportion of signal areas of the largest slice of the tumor on T 1WI and T 2WI. The classification criteria according to T 1WI: type Ⅰ, the area of hyperintensity was ≥50%; type Ⅱ, the area of hyperintensity was <50%; type Ⅲ, the tumor did not show hyperintensity, and the area of isointensity was ≥50%; type Ⅳ, the tumor did not have high signal area, and the area of low signal was ≥50%. The classification criteria according to T 2WI: type Ⅰ, the area of low signal in the tumor was ≥50%; type Ⅱ, the area of low signal was <50%; type Ⅲ, the tumor did not contain low signal area, and the area of isointensity was ≥50%; type Ⅳ, the tumor did not have low signal area, and the area of high signal intensity was ≥50%. The proportion of each type was calculated. Results:According to T 1WI, typeⅠwas identified in 27 cases (42.9%, 27/63), typeⅡ in 25 cases (39.7%, 25/63), type Ⅲ in 4 cases (6.3%, 4/63), and type Ⅳ in 7 cases (11.1%, 7/63). According to T 2WI, type Ⅰwas demonstrated in 29 cases (46.0%, 29/63), type Ⅱ in 28 cases (44.4%, 28/63), type Ⅲ in 2 cases (3.3%, 2/63), and type Ⅳ in 4 cases (6.3%, 4/63). There were 16 cases classified as type I based on T 1WI and T 2WI. Conclusions:Typical and atypical SMM can be identified according to signal patterns. The typeⅠsignal pattern of SMM cases on T 1WI and T 2WI is typical and can be easily diagnosed, but the proportion was less than 50%. For atypical SMM, malignant melanoma should be strongly suspected if hyperintense on T 1WI or hypointense on T 2WI is found.