Objective To compare effect of chemotherapy agent DDP to MACS in sorting cancer stemcells (CSC) of laryngeal carcinoma cell line Hep-2. Methods CD133 magnetic beads were applied to sort Hep-2 cells. Different dosages of DDP were used to treat Hep-2 cells for 48 hours. Enrichment rate of CD133+ cells by MACS and after DDP treatment was detected by Flow Cytometer (FCM). Morphologic change was observed under inverse-phase microscope. Results FCM showed that the sorting rate of CD133+ cells through MACS was 64.33 %, while after DDP treatment for 48 hours, the rate of CD133+ cells was enriched significantly in each dosage of DDP, with the maximal rate was 50.7 %, in the dosage of 4 μg/ml. There was a significantly difference between MACS and each of DDP group (P <0.01). Cells treated with DDP were abnormal in morphology. Conclusion MACS and DDP sorting has respective advantages in enriching CSC in Hep-2 cell lines.

Objective To analyze the clinical features and prognosis of hyperglycemia and the relationship between the blood glucose level and the severity of disease in critically ill children.Methods A total of 349 critically ill children admitted in Pediatric Intensive Care Unit (PICU) from November 2009 to April 2010 were restrospectively analyzed.According to the levels of venous blood glucose within 24 h after admission,they were divided into very high level group (blood glucose ≥11.1 mmol/L,n =67 ),slightly high level group (blood glucose 6.3-11.1 mmol/L,n =134) and normal level group (blood glucose ≤6.3mmol/L,n =148).Blood glucose levels were measured within 24 hours,3 days and 7 days after admission.Electrolytes,inflammatory markers,cardiac enzymes,liver and kidney function as well as other biomarkers related to the severity and the prognosis of the patients were recorded after admission.The categorical variables were analyzed with Chi -squared test,the continuous variables were analyzed with t-test,F-test,U-test andH-test,and the correlation analysis was calculated by using Pearson Coefficients. Results In the very high level group,slightly high level group and normal level group,the average blood glucose levels were 16.98 ±7.08 mmol/L,8.25 ± 1.40 mmol/L and 4.89 ± 0.98 mmol/L ( P ＜ 0.01 ),respectively;and the Pediatric Critical Ⅲ Scores at admission were 81.22 ± 8.25,86.71 ± 6.40 and 86.15 ± 6.99 ( P ＜0.01 ),respectively,and the incidences of sepsis or septic shock were 55.23％,30.59％ and 14.18％,respectively (P ＜0.01 ),and the incidences of MODS were 46.26％,22.39％ and 16.23％,respectively (P ＜0.01 ).The blood glucose levels of patients with one organ failure and two organ failure were 8.27 ± 3.75 mmol/L and 8.88 ± 5.42 mmol/L,respectively ( P ＜ 0.05 ).The blood glucose levels of patients with two organ failure and multiple organ failure were 8.88 ± 5.42 mmol/L and 13.09 ± 8.23 mmol/L,respectively (P＜0.01).The mortality rates of three groups were 47.76％,14.93％ and 10.13％ (P ＜0.01 ),and the blood glucose levels at admission in survival group and death group were 7.57 ±4.11 mmol/L and 12.46 ± 8.17 mmol/L ( P ＜ 0.01 ).Conclusions Patients with hyperglycemia are often found in the PICU.It not only partially reflects the severity of the disease,but also serves as an important indicator for the prognosis.The blood glucose level is positively correlated to the number of compromised organs and the severity of the disease.Dynamic monitoring of blood glucose may be essential for controlling the symptoms and prediction of prognosis.

Objective To evaluate the application of different assays for detection of human papillomavirus(HPV)in diagnosis of high grade cervical lesions.Methods Two hundred subjects with abnormal thinprep liquid-based cytology test(TCT)Resultswere selected for HPV DNA detection by hybrid capture 2(HC-Ⅱ) and Methodsbased on PCR including flow-through hybridization and gene chip (HybriMax),real-time fluorescent quantitative PCR(FQ-PCR)and flow fluorescent hybridization assay.Cytopathological Resultswere used as gold standards to evaluate the test performance of the above assays for diagnosing cervical intraepithelial neoplasia(CIN)≥Ⅱ. SPSS 13.0 software was used for statistical analysis.Results HPV DNA positive rates of 200 samples by HybriMax,FQ-PCR,flow fluorescent hybridization assay and HC-Ⅱ were 72.5%(145/200),71.5%(143/200),70.0%(140/200)and 69.0%(138/200),respectively,and the differences were not statistically si(g)nificant(x2 =0.252,0134,0.012 and 0.027,P ＞ 0.05).The sensitivity,Youden index and negative predictive value of the above assays were statistically different(x2 =7.923,7.819 and 8.108,P ＜0.05).Conclusion HC-Ⅱ is superior to PCR Methodsin diagnosis of CIN Ⅱ and above.

Objective To analyze the clinical features of the hyperamylasemia in critically ill children and investigate the related risk factors in order to provide the basis for prevention and treatment. Methods A total of 1036 critically ill children admitted in pediatric intensive care unit (PICU)from April,2011 to Oct,2012 were studied.They were divided into the high amylase group (n=82)and the normal group (n=954).According to the outcomes,the high amylase group was divided into survival group (n=61 ) and death group (n =21 ).The related risk factors of the occurrence and outcome of hyperamylasemia were analyzed by univariate and multivariate Logistic regression.Results There were statistically significant differences in rates of coagulation disorders, convulsions, disturbance of consciousness,pediatric critical illness score (PCIS)≤80,multiple organ dysfunction (MODS)≥3, sepsis,shock,and lactic acid (LA),procalcitonin (PCT),blood glucose (BG)between the high amylase group and the normal group (P<0.05 ).The differences in the rates of coagulation disorders,convulsions, mechanical ventilation,PCIS≤80,MODS≥3,and LA,PCT,oxygenation index,albumin,C-peptide,BG were statistically significant between the survival group and the death group (P <0.05 ).Multivariate Logistic regression analysis showed that the risk factors of the hyperamylasemia's occurrence were LA,PCT, BG,PCIS<80,MODS>3.Adjusted ORs confidence intervals of them were 1.662 (1.236-2.234),1.042 (1.025-1.060),1.612 (1.411-1.843),3.219 (1.311-7.905),3.411 (1.370-8.494),respectively. The hyperamylasemia's prognostic risk factors were PCT,C-peptide,PCIS ≤80,MODS >3,shock. Adjusted ORs confidence intervals of them were 1.066(1.021-1.113),1.437(1.017 ~2.030),16.137 (1.876-138.836),10.437(1.528-71.925),20.928(1.938-226.009),respectively.Conclusions The severity of the disease,the levels of LA,PCT,BG in critically ill children were positively correlated to the occurrence of hyperamylasemia.The severity of the disease,the incidence of organ failures,the levels of PCT,C-peptide combined shock in children with hyperamylasemia were positively correlated to the prognosis of hyperamylasemia.

Objective To analyze the clinical features of critically ill children with abnormal pancreatic ultrasound findings in order to explore the related risk factors for offering evidence-based diagnosis of pancreatic damage secondary to critical illness.Methods A prospective study was performed in 531 critically ill children admitted to pediatric intensive care unit(PICU) of 17 children' hospitals from January 2012 to March 2014.All patients were divided into control group(513 cases) and abnormal group(18 cases) according to the pancreatic ultrasound findings.Comparison of clinical features and biochemical indicators were made between two groups.The related risk factors associated with abnormal pancreatic ultrasound findings were analyzed by using Logistic regression analysis.ROC curves were used to evaluate the role of amylase and lipase in the diagnose of abnormal pancreatic ultrasound findings.Results The incidence of abnormal pancreatic ultrasound findings in critically ill children was 3.39%, the average age of abnormal group was significantly older than that in control group (P<0.01).There were not statistically significant differences in gender and primary disease between two groups.The incidence of hypotension was 22.2% and the incidence of abdominal muscle tension was 16.7% in abnormal group,which were statistically higher than those in control group (P<0.05).The levels of calcium, albumin of abnormal group were significantly lower than those in control group (P<0.01), and levels of serum amylase, lipase, lactate dehydrogenase in abnormal group were significantly higher than those in control group (P<0.01).The risk factors associated with abnormal pancreatic ultrasound findings were age, blood pressure, calcium,amylase.Area under the ROC curve of abnormal pancreatic ultrasound findings determined by amylase and lipase were 0.803 and 0.745,respectively (P<0.05).The sensitivity was 0.667, specificity was 0.881 when the serum amylase was 101.5 U/L, the sensitivity was 0.722, specificity was 0.928 when the serum lipase was 96.9 U/L.Conclusions The incidence of abnormal pancreatic ultrasound findings was rather low in critically ill children.The risk factors associated with abnormal pancreatic ultrasound findings were hypotension, hypocalcemia, and hyperamylasemia.The elevated serum amylase and lipase might be the most likely factors associtaed with abnormal pancreatic ultrasound findings.

OBJECTIVE To study the expression of matrix metalloproteinase 2 (MMP-2) and its correlation with microvascular density(MVD) and tumor metastasis in nasopharyngeal carcinoma. METHODS The expression of MMP-2 and MVD were detected in the specimens of 50 cases with nasopharyngeal carcinoma and 15 cases with nasopharyngeal inflammation by immunohistochemical technique. RESULTS The MVD in nasopharyngeal carcinoma(21.92?7.80) was significantly higher than that in inflammation nasopharyngeal tissues (9.23? 1.84, P

Objective Previous study found TGF-βpathway might be the molecular pathway influencing the prognosis of colo-rectal cancer, while it was uncertain whether Chinese population is associated with the disease.The article was to evaluate the genetic factors associated with prognosis in colorectal cancer. Methods 52 cases patients with colorectal cancer were followed-up for 36 months in our hospitals from January 2013 to August 2014.Their DNAs were extracted and stored and gene typing were carried out in 5 candidate genes to detect the association between SNPs and the prognosis in colorectal cancer. Results The results showed that within the TGF-βsignaling pathway, after adjusting for Bonferroni multiple testing, allele A of SNP rs10749971 located in gene POU2AF1 was associated with the recurrence of patients with stage III disease under additive and recessive genetic models ( HR =1.968, P=0.004;HR=2.174, P=0.010).Allele C of SNP rs961253 in the gene BMP2 could increase the recurrence risk (HR=1.992, P=0.005) and the death risk (HR=3.161, P=0.007) of patients with stage III disease under recessive genetic models.Allele A of SNP rs4464148 in SMAD7 gene could significantly decrease the death risk of patients with stage II and III colorectal cancer under dominant genetic model (HR=0.382, P=0.017;HR=0.230, P=0.006).In addition, accumulated effects of several adverse genes showed gene high risk group could increase the risk of death for patients with stage III colorectal cancer significantly ( HR=15.512, P=0.036;95%CI:1.611-149.360). Conclusion In different genetic models, SNP locus mutation within gene POU2AF1, BMP2 and SMAD7 on TGF-βpathway was associated with the prognosis of patients with colorectal cancer.With the increase of the number of unfavorable genes, the death risk increases accordingly.

AIM: To study the effect of hydrogen sulfide on wound healing of skin ulcer in diabetic rats . METHODS:Male SD rats were randomly divided into 3 groups, including non-diabetic control (NDC) group, untreated diabetic control ( UDC) group, and treated diabetic administration ( TDA) group.Diabetic rats were induced by intraper-itoneal injection of streptozotocin (STZ).After 1 week, wound healing model was prepared by making a round incision ( 2.0 cm in diameter ) on the dorsal skin in full thickness .The rats from TDA group received 2%sodium bisulfide ointment on the skin ulcer wound , and the animals from UDC and NDC groups received control cream .After 21 d of treatment with sodium bisulfide , blood samples were collected for biochemical analysis , including prothrombin time ( PT) , thrombin time (TT), and fibrinogen (FIB) in plasma, as well as the activity of superoxide dismutase (SOD) and the content of malondi-aldehyde ( MDA) in the serum.White blood cells ( WBC) and lymphocytes were also counted .Granulation tissues from the wound were processed for histological examination and Western blot analysis was used to detect heme oxygenase -1 ( HO-1) and tumor necrosis factor α(TNF-α) expression.RESULTS:Compared with UDC group, sodium bisulfide treatment accelerated wound healing of skin ulcer (P<0.01), and increased the activity of SOD in serum (P<0.01) in the diabet-ic rats.The declined number of WBC and lymphocytes , prolonged PT and TT , and decreased FIB levels in rats treated with sodium bisulfied were also confirmed .Pathological section showed that there were inflammatory cell infiltration , and irregu-lar and loose fibril alignment in the granulation tissue of rats from the UDC group , but there were regular fibril alignment and increased angiogenesis in the granulation tissue of rats from the TDA group (P<0.05).Furthermore, sodium bisulfide treatment raised HO-1 protein expression , and decreased TNF-αprotein expression in the diabetic rats .CONCLUSION:Hydrogen sulfide accelerates the wound healing of skin ulcer in the rats with diabetes .The beneficial effect of H 2 S may be associated with formation of granulation , anti-inflammation , and antioxidation .

ObjectiveTo explore the value of elevated amylase in assessment of severity of the disease and its inlfuencing factors in critically ill children.MethodThe clinical data from critically ill children hospitalized in pediatric intensive care unit were retrospectively analyzed from November 2009 to June 2014. According to levels of serum amylase, the critically ill children were divided into normal serum amylase group (≤103 IU/L) and elevated serum amylase group (>103 IU/L). The differences between two groups were compared.ResultsA total of 1920 critically ill children were enrolled, most of whom had primary respiratory and neurological diseases. Among them, 1470 children had normal serum amylase (76.6%) and 450 children had ele-vated serum amylase (23.4%). The elevated serum amylase group had signiifcantly higher rate of organs failure (≥4), lower rate of PCIS (<70), higher rate of invasive mechanical ventilation and mortality than those in normal serum amylase group (P<0.001). The correlation of serum amylase with lipase blood, urine amylase, blood urea nitrogen and creatinine were signiifcant (r=0.246 to 0.683,P<0.001). Blood amylase was positively correlated with shock index (r=0.111,P=0.002) and negatively correlated with respiratory failure index (r=0.133,P<0.001).ConclusionsSerum amylase could be used to assess the severity of disease. The elevated amylase was closely related to pancreatic exocrine function, renal function, ischemia, and hypoxia. The elevated serum amylase in critically ill children is a warning sign.

Nanobody is a kind of antibody from camel, which misses light chain. Nanobody has the same antigen binding specificity and affinity as mAb. Moreover, because of its small molecular weight, high stability and easy preparation, nanobody has great value of biomedical applications. In this study, we successfully prepared highly pure antiEGFR nanobody in E.coli using genetic engineering techniques. Cell proliferation assay (CCK-8 assay) and migration experiments (cell scratch test and Transwell assay) indicated that the recombinant antiEGFRnano can significantly inhibit the proliferation and migration of endometrial cancer cells. These results provide a new way of thinking and methods for EGFR-targeted therapy of endometrial cancer.