Objective To investigate the effects of miR-100 on the proliferation of MIA PaCa-2 and CFPAC-1 cells through targeting fibroblast growth factor receptor 3 (FGFR3).Methods miR-100 expression levels in 17 cancer tissues and 17 nonmalignant tissues were examined by Real-time PCR.The effect of miR-100 overexpression on cell proliferation was examined by CCK-8 assay in vitro.Luciferase assay was used to confirm that miR-100 could directly target FGFR3.Real-time PCR and Western blot were used to examine the expression of FGFR3 in miR-100 overexpressing pancreatic cancer cells.The predicted target gene of miR-100,FGFR3,was downregulated by siRNA,and its effect on cell proliferation was also examined.Cell proliferation was analyzed using CCK-8 and Edu assay.Results miR-100 was lowly expressed in pancreatic cancer tissues (P ＜ 0.05).In pancreatic cancer cells,the transfection of lv-miR-100 was able to upregulate the endogenous expression of miR-100 and inhibit the cell proliferation (P ＜0.05).Luciferase assay showed FGFR3 was the direct target of miR-1O0.FGFR3 was significantly downregulated by overexpressing miR-100 in pancreatic cancer cells (P ＜0.05),and FGFR3 knockdown by specific siRNA exerted the similar effect as miR-100 overexpression (P ＜ 0.05).Conclusions Our study identified a new miRNA regulator,miR-100,and clarified a novel mechanism of how miR-100 regulates cell proliferation in pancreatic cancer.The strategy of overexpressing the tumor suppressor miR-100 may provide a new therapeutic approach for treating patients with pancreatic cancer.

Objective: To study the relationship between Framingham risk score for coronary artery disease (CAD) and cognitive function in healthy community elders.
Methods: A total of 276 healthy community elders were evaluated by Framingham score to predict the risk for suffering from CAD in 10 years. The subjects were divided into 3 groups. High risk group (the risk > 20%), n=46, Mid risk group (the risk at 10%-20%), n=76 and Low risk group (the risk < 10%), n=154. The cognitive function was measured by mini-mental state examination (MMSE) and China adult intelligence scale (CISA). The differences of cognitive function levels to 3 CAD risk groups were studied.
Results: With the increased CAD incidence from Low risk, Mid risk to High risk groups, the MMSE score reduced accordingly (26.9 ± 1.45) vs (24.3 ± 1.53) vs (22.2 ± 1.43), P=0.014. Pearson analysis presented that MMSE score was negatively related to Framingham risk score (r=-0.213, P<0.001). There were several elements of cognitive function related to Framingham risk score including MMSE score, question answering, grid filling, oral arithmetic and word distinguishing (r=-0.247), (r=-0.167), (r=-0.132), (r=-0.152) and (r-0.256), all P<0.05.
Conclusion: CAD risk level was negatively related to cognitive function, the higher Framingham risk score resulted in the lower cognitive function in healthy community elder subjects.

BACKGROUND:Our previous studies have shown that strontium-doped calcium polyphosphate containing low-dose strontium appears to have a significant effect on angiogenesis-related behaviors of monocultured umbilical vein endothelial cells and osteoblasts.
OBJECTIVE:To investigate the effect of strontium-doped calcium polyphosphate on angiogenesis-related behaviors of umbilical vein endothelial cells and osteoblasts co-cultured, including celladhesion, spreading, proliferation, as wel as the protein secretion of vascular endothelial growth factor and basic fibroblast growth factor from co-culture system in vitro.
METHODS:Human umbilical vein endothelial cells and osteoblastic cells (MG63) were utilized in this study. cells from passage 3 were used for preparation of the cel-scaffold constructs. After placed in 24-wel plate at a ratio of 2:1, human umbilical vein endothelial cells and MG63 cells were seeded onto strontium-doped calcium
polyphosphate, calcium polyphosphate and hydroxyapatite scaffolds and co-cultured for 7 days. The vascular endothelial growth factor and basic fibroblast growth factor protein levels were determined through a double ligand enzyme-linked immunosorbent assay. The colorimetric 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay was performed to quantify the effect of scaffolds on cellproliferation.
RESULTS AND CONCLUSION:Compared with those on calcium polyphosphate and hydroxyapatite scaffolds, cells on strontium-doped calcium polyphosphate scaffolds attached and spread better with a significantly improved cellproliferation. More importantly, the vascular endothelial growth factor and basic fibroblast growth factor expressions were significantly higher in the strontium-doped calcium polyphosphate group than the other two groups (P<0.05), indicating strontium-doped calcium polyphosphate can up-regulate levels of vascular endothelial growth factor and basic fibroblast growth factor proteins.

Objective To observe the effects of modified Jianpi Yishen Decoction on urinary osteopontin (OPN) in calcium oxalate nephrolithiasis patients after percutaneous nephrolithotomy (PCNL) or ureteroscope lithotomy (URL);To clarify the mechanism of modified Jianpi Yishen Decoction on the prevention of calcium oxalate kidney stones. Methods Totally 116 calcium oxalate nephrolithiasis patients were randomly divided into trial group (62 cases) and control group (54 cases). The trial group took modified Jianpi Yishen Decoction every other day, while the control group took potassium sodium hydrogen citrate granules three times a day. The concentrations of OPN, urinary calcium and urinary oxalic acid of the patients in the two groups were observed before treatment and 2 weeks and 4 weeks of treatment. Results The concentration of urinary OPN of 2 weeks and 4 weeks of the treatment in the trial group was significantly increased compared with before treatment (P<0.05), and the difference was statistically significant compared with the control group (P<0.05). The concentration of urinary OPN in the control group had no significant change after treatment (P>0.05). The differences in the concentrations of urinary calcium and urinary oxalic acid of the two groups between before and after treatment were not significant (P>0.05). Conclusion Modified Jianpi Yishen Decoction can effectively restrain the formation of the calcium oxalate stones by increasing the level of urinary OPN, which demonstrates effective prevention in the calcium oxalate nephrolithiasis patients after PCNL or URL.

Objective To observe the incidence and the influencing factors of kidney insufficiency in acute cerebral infarction patients with non-valvular atrial fibrillation. Methods From January 2013 to January 2015,266 consecutive acute cerebral infarction patients with non-valvular atrial fibrillation admitted to the Department of Neurology,Xuanwu Hospital,Capital Medical University were enrolled retrospectively. Renal function was assessed by the estimated glomerular filtration rate (eGFR),eGFR <60ml/(min·1.73 m2 ) was defined as renal insufficiency,and were divided into a renal insufficiency group (n = 36)and a non-renal insufficiency group (n = 230). The incidence of kidney insufficiency and its influencing factors in acute cerebral infarction patients with non-valvular atrial fibrillation were observed. Results (1)In 266 acute cerebral infarction patients with non-valvular atrial fibrillation,the prevalence of renal insufficiency was 13. 5% (n = 36). The proportion of age (≥65 years)of the renal insufficiency group was higher than that of the non-renal insufficiency group. There was significant difference (94. 4%[34 / 36]vs. 70. 0%[161 / 230];P = 0. 002). There were no significantly differences in general information of others (all P >0. 05). (2)Multiple Logistic regression analysis showed that the age (≥65 years)was an independent risk factor for the occurrence of renal insufficiency in acute cerebral infarction patients with atrial fibrillation (OR,1. 147,95% CI 1. 087 -1. 209;P < 0. 01),and the histories of hypertension (OR,0. 870,95% CI 0. 362-2. 089;P = 0. 755),diabetes mellitus (OR,1. 078,95% CI 0. 403 -2. 883;P = 0. 882 ), and hyperlipidemia (OR,1. 666,95% CI 0. 645 - 4. 302;P = 0. 292 )were not associated with renal insufficiency in cerebral infarction patients with atrial fibrillation. Conclusions The incidence of renal insufficiency in cerebral infarction patients with atrial fibrillation is higher. Age (≥65 years)is an independent risk factor for renal insufficiency in this type of patients.

Objective To investigate the protective effects of Ulinastatin on intestinal barrier damaged after cardiopulmonary resuscitation (CPR) in rats in order to illustrate the possible mechanism.Methods Twenty-one male SD rats were divided into three groups randomly (random number) including control group (sham group, n =7), cardiopulmonary resuscitation group (CPR group, n =7) and ulinastatin group (UTI group, n =7).The rats were anesthetized with pentobarbital sodium (45-60 mg/kg) by intraperitoneal injection.The rats of sham group were only treated with endotracheal intubation.Ulinastatin (100 000 U/kg) were injected via caudal vein 2 hours prior to CPR, and cardiac arrest was made in rats and cardiopulmonary resuscitation was carried out in the UTI group, while equivalent volume of sterile saline was used instead in the CPR group.Blood and ileum samples were obtained at 48 hour after restoration of spontaneous circulation (ROSC).The levels of tumor necrosis factor alpha (TNF-α) and interleukin-1β (IL-1β) were assayed by ELISA (enzyme-linked immunosorbent assay), the protein levels of caspase-3 were determined by western blot, the intestinal mucosa were stained by terminaldeoxynucleoitidyl transferase mediated nick end labeling (TUNEL) and ileac mucosa were observed under transmission electron microscope.Data were processed with SPSS 17.0 software.Results The plasma levels of TNF-α and IL-1β were dramatically higher in CPR group than those in other two groups (CPR vs.sham, P ＜ 0.01;CPR vs.UTI, P ＜ 0.05).Moreover, the tight junctions between cells obviously broadened and loosened in the CPR group were found under electron microscope, however, this phenomenon was not obvious in the UTI group.A large number of apoptotic cells were observed by TUNEL assay in the CPR group, but a small number of apoptotic cells were observed in the UTI group.The protein levels of caspase-3 in the UTI group were higher than those in sham group, but lower than those in CPR group (both P ＜ 0.05).Conclusions Ulinastatin has protective effects on the intestinal barrier damaged after cardiopulmonary resuscitation in rats by decreasing the proinflammatory mediators in the blood, reducing the expression of caspase-3and then reducing the numbers of apoptotic intestinal cells.

Objective To investigate the effects and mechanism of expression of microRNA-100 in the tissues of pancreatic cancer and its relationship with clinicopathological factor.Methods The clinical data of 17 patients with pancreatic cancer who were admitted to the Affiliated Hospital of Guizhou Medical University between January 2013 and March 2014 were retrospectively analyzed.The surgical specimens were collected for study.The expression of microRNA-100 in the pancreatic tissues were detected by real-time fluorescent quantitative polymerase chain reaction (RT-PCR) and its relationship with clinicopathological factors was analyzed.The MIA PaCa-2 cells and CFPAC-1 cells were infected by lv-microRNA-100.MIA PaCa-2 cells were divided into the M group (microRNA-100 were infected) and N group (empty vectors were infected),and CFPAC-1 cells were divided into the C group (microRNA-100 were infected) and D group (empty vectors were infected).The expressions of microRNA-100 in the CFPAC-1 cells and MIA PaCa-2 cells were detected by RT-PCR and cell cycles were detected by flow cytometry.The expressions of Cyclin D1 protein in the CFPAC-1 cells and MIA PaCa-2 cells were detected by Western blot.Measurement data with normal distribution were presented as (x) ± s and comparison among groups were done by t test.Results The results of RT-PCR showed that the relative quantitative expression of microRNA-100 in the pancreatic cancer tissues was 0.046 ± 0.020,which was significantly different from 0.097 ± 0.017 in the adjacent tissues (t =2.789,P ＜ 0.05).There were significant differences between the tumor differentiation degree and tumor staging in patients with pancreatic cancer (t =2.563,2.135,P ＜0.05).The results of RT-PCR showed that the relative quantitative expression of microRNA-100 in the M group was 0.097 ±0.012,which was significantly higher than 0.018 ± 0.006 in the N group (t =4.410,P ＜ 0.05).The relative quantitative expression of microRNA-100 in the C group was 0.084 ± 0.021,which was significantly higher than 0.023 ± 0.010 in the D group (t =5.351,P ＜ 0.05).The results of flow cytometry showed that the percentage of cells between G0-G1 were 45.3％ ±0.7％ in the M group and 30.6％ ±0.7％ in the N group,with a significant difference (t =5.564,P ＜ 0.05).The percentage of cells between G0-G1 were 58.8％ ± 1.0％ in the C group and 42.6％ ± 0.7％ in the D group,with a significant difference (t =7.771,P ＜ 0.05).The results of Western blot showed that the relative quantitative expression of Cyclin D1 protein were 0.352 ± 0.081 in the M group and 0.872 ± 0.134 in the N group,with a significant difference (t =7.651,P ＜ 0.05).The relative quantitative expression of Cyclin D1 protein was 0.410 ± 0.121 in the C group,which was significantly different from 0.979 ± 0.232 in the D group (t =8.712,P ＜ 0.05).Conclusion Low expression of microRNA-100 in pancreatic cancer tissues may be correlated with tumor differentiation degree and tumor staging,it can inhibit tumor cells proliferation by reducing expression of Cyclin D1 protein.

OBJECTIVE:To screen and identify endophytic fungi from Schisandra chinensis with antioxidant activity. METH-ODS:The tissue isolation skill was used to isolate endophytic fungi from roots,leaves,stems and fruits of S. chinensis. And anti-oxidant activity of endophytic fungi were screened by DPPH radical scavenging assay and hydroxyl radical scavenging assay. The to-tal DNA were extracted;the 18S rDNA ITS were amplified and sequenced with primer ITS1 and ITS4;the results of sequencing were analyzed comparatively based on homology to confirm the classification of active strains. RESULTS:23 strains were isolated from S. chinensis. GSR-12,isolated from roots of S. chinensis,had strong antioxidant activities. The scavenging rate on DPPH and the hydroxyl radical were 87.96% and 82.31% respectively. GSR-12 strain was identified as Clonostachys rosea by analyzed com-paratively. CONCLUSIONS:1 strain of C. rosea,isolated from roots of S. chinensis,has strong antioxidant activity.

Objective To investigate the expression profiles of microRNA in early esophageal squamous cancer and normal esophageal tissues and verify the significantly different expression miRNAs , further to study the effects on proliferation of EC109. Methods The microarray assay was performed to analyze miRNA expression profiles in three pairs of early esophageal squamous cancer and the corresponding normal esophageal tissues. Quantitative real-time polymerase chain reaction (qRT-PCR) was used in another 38 pairs samples to further verify the differentially expressed miRNAs . Three verified miRNAs ( miR-29a, miR-221 and miR-222) mimics were transfected into EC109 respectively and CCK8 method was used to study the effect of cell proliferation in each miRNA. Results Microarray technique selected 53 miRNAs that differentially expressed in early esophageal squamous cancer and normal esophageal tissues , 32 miRNAs were up-regulated and 21 miRNAs were down-regulated. qRT-PCR verified that miR-29a was significantly down-regulated (P < 0.05) and miR-221, miR-222 were significantly up-regulated (P < 0.05) in early esophageal squamous cancer tissue. Over-expression of miR-29a could significantly inhibit the proliferation of EC109 (P < 0.05) whereas over-expression of miR-221 or miR-222 could both significantly promote the proliferation of EC109 (P < 0.05). Conclusion There was significant difference of miRNAs expression between early esophageal squamous cancer and normal esophageal tissues, and the differentially expressed miRNAs could be used as new biomarkers for early diagnosis of esophageal squamous cancer.

Objective To explore the relationship between the rs18004197 polymorphism in the 3'-untranslated region of adenomatous polyposis coli (APC) gene and colorectal cancer susceptibility.Methods Firstly,we collected the peripheral venous blood of 573 colorectal cancer cases and 588 controls,and then extracted DNA from blood samples,genotyped rs1804197 polymorphism using real-time PCR and assessed its association with the susceptibility of colorectal cancer.Results There were 387 CC (67.5％),153 AC (26.7％) and 33 AA (5.8％) genotypes in the colorectal cancer cases.In the control group,there were 427 CC (72.6％),144 AC (24.5％) and 17 AA (2.9％) genotypes.The AA genotype odds ratio (OR =2.14,95％ CI:1.17-3.91,P =0.011) and the A allele frequency (P =0.011) were significant difference in case and control groups.Further subgroup analysis showed that the differences of the frequency distribution in the male (P =0.048) and non-drinking (P =0.020) groups were statistically significant.In the male group,the risk of colorectal cancer was increased by 0.41 (OR =1.41,95％ CI:1.01-1.98) for individuals bearing the A allele.In the non-drinking group,the risk of colorectal cancer was increased by 0.22 (OR =1.22,95％ CI:0.91-1.64) for individuals bearing the A allele,but the result was not statistically significant.Conclusion The rs18004197 polymorphism in the 3'-untranslated region of APC gene is related to the susceptibility of colorectal cancer.The AA genotype may increase the susceptibility of colorectal cancer.