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 investigate the incidence and influencing factors for sharp injuries occurred in workers in central sterile supply departments (CSSDs).Methods In October-November 2012,a multicenter cross-sectional survey was conducted,a total of 95 workers in CSSDs of 16 hospitals in Hunan Province were surveyed through questionnaires.Results A total of 89 workers (93.68%)in CSSDs sustained sharp injuries,46 (48.42%)of whom sustained 1 -5 times of sharp injuries,22(23.16%)sustained 6-10 times of sharp injuries,and 11 (11 .58%)sus-tained for at least 10 times of sharp injuries.81 workers (85.26%)sustained at least one time of sharp injuries one year before survey,52 (54.73%)of whom were injured by contaminated needles or other sharps.Logistic regression analysis revealed that higher educational background was protective factor for sharp injuries(OR 90%CI :0.05-0.87);while poor sleep quality(OR 90%CI :1 .03-17.94),frequent touching sharps(OR 90%CI :1 .11 -12.15),and irrational placing of objects by the other medical stuff (surgeons and nurses in operating rooms)(OR 90%CI :1 .23-16.98)were the risk factors for sharp injuries.Conclusion The incidence of sharp injuries among workers in CSSDs is high,which is related to personal factors and environmental factors.It is suggested to strengthen staff training to enhance their awareness of precaution.

Objective To synthesize 18F labeled N-(2-18 F-fluoropropionyl)-L-glutamine (18 F-FPGLN) for tumor PET imaging,and to perform its biodistribution study on normal mice and PC-3 tumorbearing nude mice.Methods 4-nitrophenyl-2-18F-fluoropropionate (18F-NFP) was synthesized on the MF2V-IT-I synthesizer and was purified by semi-preparative HPLC.Anhydrous 18F-NFP was added to a solution of L-glutanine t-butyl ester to synthesize 18F-FPGLN t-butyl ester,which was hydrolyzed by HCl (3 mol/L) and neutralized with NaOH (2 mol/L) solution.18F-FPGLN product was collected for further study.Biodistribution study was performed on normal Kunming mice and PC-3 prostate cancer tumor-bearing nude mice,respectively.Results 18F-FPGLN was synthesized with 10％-15％ (decay uncorrected) overall radiochemical yield after 130 min of radiosynthesis.The radiochemical purity was higher than 96％.Rapid and high uptake of radiotracer was observed within the kidneys,and was quickly excreted through the urinary bladder.The uptake in kidney reached (35.0±1.2) ％ID/g at 5 min post-injection,and descended to (1.5±0.3) ％ID/g at 120 min.The liver,lung,heart and small intestine showed relatively moderate uptake of radioactivity.The uptake in the pancreas,muscle,spleen,stomach and brain was low,and the lowest uptake of (1.5±0.3) ％ID/g was found in the brain at 30 min post injection.High accumulation of 18F-FPGLN in PC-3 xenograft was observed,and the tumor/muscle ratio reached 2.07 at 60 min post injection.Conclusion A novel N-position 18F-labeled glutamine analogs 18F-FPGLN,with favorable pharmacokinetic characteristics,is synthesized successfully,which makes it possible to perform tumor PET imaging using 18F-FPGLN subsequently.

Objective To investigate the application of statins in secondary prevention of ischemic stroke and transient ischemic attacks (TIA) in different risk groups,and to identify the factors influencing the compliance of statins. Methods Data were prospectively collected on consecutively encountered ischemic stroke or TIA patients admitted to the First Affiliated Hospital of Zhengzhou University from April 2009 to January 2010.All clinical characteristics and possible factors influencing the compliance of statins were collected; the application of statins was investigated at 3-month follow-up.The multivariate Logistic regression analysis was used for the analysis of influence factors of the compliance of statins.Results All 369 patients were collected,52.8％ of cases were prescribed statins for therapy during hospitalization.The application rate of statins in accordance with guidelines in high-risk group,extremely high-risk Ⅱ group and extremely high-risk Ⅰ group was 25.0％ (16/64),44.1％ (30/68) and 71.4％ (135/189),respectively. Logistic regression analysis showed that the statins application during hospitalization was associated with diabetes history ( P =0.032,OR =1.789,95％ CI 1.052-3.043 ) and the presence of carotid vulnerable plaques(P =0.000,OR =5.308,95％ CI 3.340-8.434).The general application rate of statins was 22.3％ (81/363),which was significantly lower than that during hospitalization. The application rate of statins in accordance with guidelines in high-risk group,extremely high-risk Ⅱ group and extremely high-risk Ⅰ group was 9.7％ ( 6/62 ),25.8％ (17/66) and 29.4％ (55/187) respectively.Logistic regression analysis showed that good compliance of statins was associated with discharge instructions on statins application ( P =0.000,OR =34.852,95％ CI 14.673-175.452 ). Conclusions The compliance of statins in secondary prevention of ischemic stroke and TIA is poor,and there is still a large gap between clinical practice and guidelines; Discharge instructions on statins application increase the compliance of statins.

Objective To investigate the predictive value of the modified National Institutes of Health Stroke Scale ( mNIHSS),measured within 9 days after stroke,for the outcome in terms of modified Rankin Scale (mRS) at 6 months after stroke.Methods AII 161 patients with acute anterior circulation ischemic stroke were recruited consecutively from July 2010 until November 2010.The mNIHSS score was assessed within 3 days,4 to 6 days and 7 to 9 days after stroke,and the mRS score at 6 months after stroke was assessed by a neurology resident.Spearman rank correlation and ROC curve were used for statistic analysis.Results One hundred and sixty-one patients were assessed within 3 days,at 4 to 6 days and 7 to 9 days after stroke.Significant Spearman rank correlation coefficients were found between mRS at 6 months and mNIHSS scores within 3 days (rs =0.592,P＜0.01 ),4 to 6 days (rs =0.597,P＜0.01 ) and 7 to 9days (rs =0.595,P＜0.01).At 6 months after stroke,the AUC ranged from 0.789 (95％ CI 0.708-0.856) for measurements within 3 days to 0.792 (95％ CI 0.712-0.857 ) and 0.799 ( 95％ CI 0.721-0.864) for 4 to 6 days and 7 to 9 days,respectively. There was no significant difference for the AUCbetween them.The optimal cut-off values for the prediction of prognosis at 6 months were 8,7 and 6 points of mNIHSS score within 3 days,4 to 6 days and 7 to 9 days,respectively.The negative predictive value gradually increased from 0.847 for assessment within 3 days to 0.867 ( 95％ CI 0.578-0.765 ) for 7 to 9 days,whereas positive predictive value declined from 0.601 for assessment within 3 days to 0.533 for 7 to 9 days.The overall accuracy of predtictions decreased from 0.753 for assessment within 3 days to 0.709 for 7 to 9 days.Conclusions When measured within 9 days,the mNIHSS has a good predictive value for final outcome in terms of mRS at 6 months after stroke.However,the optimal cut-off values for the prediction of prognosis at 6 months are different when early mNIHSS scores were assessed at different periods.

Objective:This study aimed to investigate the possible mechanism of 32P colloid induced apoptosis of craniopharyngi-oma (CP) cells in vitro and the relationship between dose effect and time effect. Methods:This study established a primary cell culture of CP limited subculture cell line. Methyl thiazolyl tetrazolium (MTT) assay was performed to plot the cell survival curve after the CP cells were treated with 32P colloid at different concentrations and time. Apoptotic rate was detected by flow cytometry(FCM). Apoptosis related DNA was investigated by TUNEL fluorescent staining. The morphological characteristics of apoptotic cells were determined by Hoechst33342 fluorescence staining. The ultrastructure of apoptotic cells was investigated by transmission electron microscopy (TEM). Results:Hoechst33342 fluorescence staining, TUNEL fluorescence staining, and TEM revealed that 32P colloid induced the apoptosis of CP cells. 32P colloid reduced the survival rate and increased the apoptotic rate of CP cells as concentration (0 MBq/mL to 14.80 MBq/mL) and time (1 d to 14 d) were increased. Conclusion: 32P colloid could effectively inhibit the growth of CP cells and induce apoptosis in vitro. High concentrations and prolonged time could induce a remarkable effect.

Objective To investigate the potential role of Spa-1 in the development and metastasis of EMPD.Methods Tissue specimens were resected from 17 patients with primary EMPD,12 patients with invasive EMPD and 9 normal human controls.Immunohistochemistry was performed to measure the expression of spa-1 and Ki-67.Results Positive staining was observed for Spa-1 in cytoplasm,and for Ki-67 in cell nuclei.Spa-1 and Ki-67 were weakly expressed in the basal layer of normal skin.The expression of Spa-1 and Ki-67 in EMPD tissue were statistically higher than those in the normal control tissue (both P<0.01).Increased expression of Spa-1 was noted in invasive EMPD tissue compared with in situ EMPD tissue.The expression of Spa-1 was positively correlated with that of Ki-67 in the tissue of EMPD.Conclusion Spa-1 may play a certain role in the initiation and progression of EMPD.

Purpose To investigate the role of SOCS3 on diabetic renal injury. Methods Male CD-1 mice were randomly divided into four groups:control group, diabetic group, empty plasmid vector transfection group and SOCS3 plasmid transfection group. The diabet-ic mice were induced by intraperitoneal injection of STZ at a dose of 150 mg/kg body weight. The mice of transfection group were re-ceived an injection of SOCS3 plasmid or empty vector at every 7 days thereafter. Specimens were collected at 12 week after STZ injec-tion. The morphological changes of tubular epithelial cells were observed by transmission electron microscope. RT-PCR and immuno-histochemistry were used to determine the mRNA and protein expression of CK18 and α-SMA. Western blotting analysis was used to determine the protein expression of SOCS3, p-STAT3, CK18 and α-SMA. Results SOCS3 overexpression in kidney down-regulated the levels of p-STAT3 andα-SMA but up-regulated the expression of CK18. Conclusion Overexpression of SOCS3 can ameliorate the tubular epithelial-mesenchymal transdifferentiation of diabetic mice via inhibiting the phosphorylation of STAT3.

BACKGROUND: Inhibiting the degradation of extracellular matrix in the intervertebral disc can delay the degenerative process of intervertebral disc. Matrix metalloproteinases-3 (MMP3) is considered as a key enzyme for degradation of extracelular matrix components such as type II collagen and aggrecan.
OBJECTIVE:To construct the short hairpin RNA lentiviral vector targeting human MMP3 gene and to detect its efficiency of gene silence by infecting human degenerative nucleus pulposus cells.
METHODS:According to the human MMP3 mRNA (NM_002422.4) sequence, four groups of the short hairpin RNA gene sequences targeting MMP3 were designed, synthesized and annealed to form double stranded DNA fragments, which were connected with the LV3 vectors digested by BamHI andEcoRI enzymes, and then transfected into the competent cels. The positive clones were identified by PCR, and analyzed by sequencing. The packaging and titer of lentivirus were determined after transfecting 293T cells. Human degenerative nucleus pulposus cels were infected with lentivirus vector, and the transfection efficiency of each group was observed under inverted fluorescence microscope. The interfering efficiency was detected by real time-PCR and western blot at 72 and 96 hours.
RESULTS AND CONCLUSION:The ds-oligo DNA was successfully inserted into the lentiviral vector as confirmed by electrophoresis and sequence analysis. The recombinant lentivirus was harvested from 293T cels with a viral titer of 1-5 ×108 TU/mL. RNA interference targeting the GCC AGG CTT TCC CAA GCA AAT sequences with the highest interfering efficiency in MMP3 gene at 72 and 96 hours resulted in suppression of MMP3 mRNA expression by 98% and 72%, respectively; and at 96 hours, the interfering efficiency of protein expression was 57.2%. The recombinant lentivirus vector containing RNA interference targeting MMP3 gene is successfuly constructed, which lays a foundation for further studies on the MMP3 function and gene therapy.

Objective:A large single-center, premature acute myocardial infarction (AMI) age (≤45 years) cohort was established to investigate the clinical features and the factors affecting major adverse cardiac events (MACE).Methods:This is a prospective and observational study. 603 patients with a clear diagnosis of AMI admitted to the Tianjin Chest Hospital from March 2015 to December 2017 were continuously selected. All patients were aged ≤45 years old, and a single-center large-sample premature AMI cohort was established. The patient's clinical basic conditions, laboratory indicators, imaging data, coronary angiography and treatment were collected. All patients were followed up for 1 year. MACE events such as cardiac death, recurrent AMI, revascularization, severe heart failure requiring hospitalization and stroke were recorded. Kaplan Meier method was used to draw the survival curve. Cox regression analysis was used to analyze the influence of risk factors, clinical characteristics and intervention methods on the long-term prognosis of MACE events.Results:A total of 603 AMI patients were included, 575 males (95.36%), 28 females (4.64%), and median age 41 (37, 44) years old. There were 422 patients (69.98%) with acute ST segment elevation myocardial infarction (STEMI), 206 patients (48.82%) with anterior myocardial infarction, and 181 patients (30.02%) with non ST segment elevation myocardial infarction (NSTEMI). Smoking was the most common risk factor for premature AMI (77.45%), followed by hyperlipidemia (48.42%) and hypertension (48.09%); smoking was the most common risk factor for male patients (80.35%), and hyperlipidemia was the most common risk factor for female patients (35.71%). 302 (50.08%) patients with premature AMI were treated with symptom onset to first medical contact (SO-to-FMC) ≤12 h; 563 patients (93.37%) had coronary angiography; coronary angiography showed that no significant stenosis, single-vessel disease, double-vessel disease, three-vessel disease, and patients with left main disease were 15(2.66%), 212(37.66%), 153(25.37%), 167(29.66%), 16(2.84%) cases; 318(56.48%) patients with vascular occlusion; The proportion of male combined with left main lesions was lower than that of female group (2.41% vs 12.50%, P=0.026); A total of 45 patients (7.46%) were recorded MACE. The 1-year MACE incidence was lower in the male group than in the female group (6.96% vs 17.86%, P=0.032). Multivariate COX regression analysis: there were 5 indicators that entered the regression model and were statistically significant: female ( HR:4.184; 95% CI:1.583-11.064; P=0.004), SO-to-FMC≤12 h ( HR:0.447; 95% CI:0.224-0.889; P=0.022), left ventricular ejection fraction (LVEF)≤40% ( HR:3.727; 95% CI:1.876-7.405; P<0.001), low-density lipoprotein (LDL) ( HR:1.315; 95% CI:1.041-1.662; P=0.022), homocysteine (Hcy) ( HR:1.011; 95% CI:1.002-1.019; P=0.011) were independent predictor of MACE occurrence in patients with early-onset AMI within 1 year. Conclusions:Smoking is the most common risk factor for young men with AMI. The most common risk factors for young women's AMI is hyperlipidemia, and the proportion of patients with left main artery disease is higher than that of men, but the proportion of patients receiving emergency intervention is lower than that of men, and the long-term prognosis of young women is poor. Early detection and control of these risk factors is a key measure to prevent the onset of AMI.