Objective To study the effect of adeno associated virus hepatocyte growth factor K1(AAV‐HGFK1)on the prolifer‐ation of 4 different colorectal cell lines with or without KRAS or BRAF mutation .Methods The levels of epidermal growth factor receptor (EGFR) mRNA were determined in SW48 without KRAS or BRAF mutation ,Lovo with KRAS mutation ,SW620 with KRAS mutation ,HT29 with BRAF mutation by quantitative real time PCR ,respectively .After the infection of AAV‐HGFK1 ,the expressions of EGFR ,p‐EGFR and β‐actin were detected by Western blot and the proliferation of the cells were assayed using MTT .Results Lovo ,SW48 and HT29 expressed EGFR protein while SW620 did not .EGF promoted the proliferation of Lovo , SW48 and HT29 cells .AAV‐HGFK1 down‐regulated the phosphorylation of EGFR and significantly inhibited their proliferation . But EGF had no effect on proliferation of SW620 stimulated by EGF .Conclusion AAV‐HGFK1 exhibited its antitumor effects through EGF/EGFR signaling irrespective of the KRAS or BRAF mutation and may also act through other signaling pathways .

Objective To assess the performance of the methylation‐sensitive high‐resolution melting curve analysis (MS‐HRM analysis) on the detection of the methylation in stool DNA for colorectal cancer screening .Methods Eighty‐two qualified stool samples were collected from 27 patients with colorectal cancer patients (CRC group) ,25 patients with advanced adenomas (AA group) ,and 30 healthy people (control group) .The methylation status of vimentin gene in all of the stool samples was detected by the MS‐HRM analysis on the LightCycler 480 platform .The fecal occult blood test (FOBT) was also used for the same samples . Results The positive rates of the MS‐HRM assay in the CRC group ,AA group ,and control group were 81 .5% (22/27) ,80 .0%(20/25) ,and 6 .7% (2/30) respectively .The positive rates of FOBT in the three groups were 37 .0% (10/27) ,12 .0% (3/25) and 3 .3% (1/30) respectively .The diagnostic sensitivity of the MS‐HRM assay for colorectal cancer and advanced adenomas (80 .8% , 42/52) was significantly higher than that of FOBT (25 .0% ,13/52)(P<0 .05) .No significant difference was found in the diagnos‐tic specificity between the MS‐HRM assay (93 .3% ,28/30) and the FOBT (96 .7% ,29/30) (P>0 .05) .Conclusion MS‐HRM performs better than FOBT and has great application potential in the detection of stool DNA methylation for colorectal cancer screening .

Objective To study the expression and the mechanism of miR-155in the villi of patients with unexplained recurrent spontaneous abortion (URSA).Methods The expression of miR-155 in the villi of 36 cases with URSA (URSA group) and 25 women with normal early pregnancy (control group) were detected by stem-loop real-time reverse transcription (RT) qPCR.Expression of hypoxia inducible factor-1 (HIF-1α),vascular endothelial cell growth factor(VEGF) and micro lymphatic vessel density (MVD) in the villi of were measured by immnohistochemical staining among two groups.Results (1) miR-155 expression:the mean miR-155 expression were 1.456 (0.489,2.459) in URSA group and 2.833 (1.740,3.794) in control group,which reached statistical difference (P ＜0.05).The mean expression of miR-155 of 1.683 (0.902,2.459) in URSA group with abortion times (≤ 3) was significantly higher than 1.229 (0.489,1.719) in URSA group with more than 4 times abortion (P ＜ 0.05).(2) Indexes:the expression of HIF-1α,VEGF and MVD value were 121 ± 12,134 ± 12,36 ± 6 in URSA group and 99 ± 10,109 ± 10,28 ±4 in control group,which reached statistical difference(P ＜ 0.01).The expression of HIF-1α,VEGF and MVD value of 119 ± 12,134 ± 12,35 ± 5 in URSA group with less than 3 times abortion was significantly lower than 128 ± 12,138 ± 12,43 ± 6 in URSA group with more than 4 times abortion (P ＜ 0.01).Conclusions The expression of miR-155 and HIF-1α is topically stimulated by oxygen signal.HIF-1α adjusts the transcription and translation of VEGF,which together involved in placental trophoblast invasion and placental angiogenesis.The low expression of miR-155 could interfere with expression of HIF-1α and VEGF,which might be involved in villous vascular dysplasia in URSA.

Objective Nanoknife,also called irreversible electroporation,is a new technique of tissue ablation.Short,microsecond electrical pulses with high voltage are applied to the cell membrane,causing pores to form within the membrane and finally leading to cell death.The current study was to investigate the efficacy and safety of the nanoknife in the ablation of the healthy pig pancreas.Methods Three healthy pigs underwent open pancreatic tissue ablation with nanoknife,and blood leukocytes and amylase were detected before and after treatment.Three pigs were sacrificed and gross specimens were collected on day 5,day 10 and day 15 after the procedure,respectively.HE staining and TUNEL staining were conducted and tissue,cellular and subcellular structures were observed under the ordinary microscope and transmission electron microscopy.Results Three experimental pigs recovered well after the procedure.No significant adhesions were found surrounding the pancreatic tissue,and the ablation zone was slightly harder.Transiently increased leukocyte count and amylase level were observed after the ablation,which decreased to the normal level on day 3 after treatment.Under light microscope,the pancreatic tissues in ablation zone appeared to be significantly different from the normal surrounding regions,with more cell death and more apoptotic cells detected by TUNEL staining.The subcellular structure changes also changed under electron microscope.But the main pancreatic duct and its large branches,together with arteriovenous distributions did not change much.Conclusions Nanoknife pancreatic tissue ablation can induce irreversible damage.In the ablation area,pancreatic duct and vascular structures are kept intact.Within a reasonable voltage range and appropriate electrical pulses setting,nanoknife ablation is safe in vivo experiment.

Objective To investigate the diagnostic efficacy of fecal calprotectin(FC) on assessing endoscopic disease activity in colonic or ileo-colonic Crohn disease (CD) and CD-related surgery patients .Methods Totally 56 colonic or ileo-colonic CD pa-tients ,25 CD-related surgery patients and 25 irritable bowel syndrome (IBS) patients with previously confirmed diagnosis of CD and IBS were enrolled into this study .Fecal samples were collected from 1 to 3 day before bowel preparation and FC was measured by ELISA .Endoscopic activity was determined for colonic or ileo-colonic CD with Simple Endoscopic Score for Crohn′s Disease (SES-CD) and CD-related surgery patients with the Rutgeerts′ score .Results Among colonic or ileo-colonic CD patients ,the levels of FC in endoscopic active patients had significantly higher than that of endoscopic remission patients and IBS patients(P < 0 .01) ,there was significant correlation between levels of FC and the SES-CD (r= 0 .802 ,P< 0 .01) .FC threshold of 250 μg/g was tested to in-dicated active endoscopic disease with 97 .1% sensitivity and 71 .4% specificity .Among CD-related surgery patients ,FC level in IBS patients were significantly lower than that of endoscopic remission patients and endoscopic active patients ,but the FC in endoscopic remission patients and endoscopic active patients had no statistic difference(P> 0 .05) ,FC cutoff level of 250 μg /g gave a sensitivity and specificity of 50 .0% ,66 .7% ,respectively .Conclusion FC is a surrogate marker for the evaluation of colonic or ileo-colonic CD endoscopic disease activity .The FC ,however ,can not distinct remission period and active period after CD surgery .

Objective To study the effect of Bairui granule and Deacetylated Ligustrazin injection on plasma BNP and FABP levels in children with heart failure and pneumonia.Methods 60 children of heart failure and pneumonia who received therapy from July 2014 to June 2016 in our hospital were randomly divided into two groups, the observation group and the control group, 30 cases in each group.children in the observation group were treated with sodium phosphate creatine, and the control group was treated with 2-linolenic acid,the clinical outcomes, clinical signs, BNF, FABP, LVEF and LVEDP were compared between the two groups.Results After treatment, the total effective rate of the observation group was significantly higher than that of the control group, the difference was statistically significant (P<0.05); The heart rate, respiratory rate and blood oxygen saturation of the two groups were significantly improved compared with those before treatment ( P<0.05 ) , the heart rate and respiratory rate of the observation group were lower than those of the control group, the oxygen saturation was higher than that of the control group (P<0.05).After treatment, the BNF and FABP indexes of the two groups were lower (P<0.05), and the BNF and FABP indexes in the observation group were lower than those in the control group (P<0.05); The LVEF of the observation group was higher than that of the control group, the LVEDP index was lower than that of the control group, the difference was statistically significant (P<0.05).Conclusion Bairui granule and Deacetylated Ligustrazin injection in the treatment of heart failure with pneumonia has a good effect in the process, which will help reduce the BNP index in children, recovery of children's body function.

Objective Diterpene ginkgolides meglumine injection (DGMI) is widely used in patients with stroke, but its efficacy and safety are not consistent.We performed a Meta-analysis to comprehensively evaluate the efficacy and safety of DG-MI in acute ischemic stroke and recovered stroke.Methods The wanfang, VIP, CNKI and PubMed were searched, the randomized controlled trials (RCTs) were enrolled.Data collection and quality evaluation of the included RCTs were performed according to Cochrane systematic evaluation method.Meta-analysis was performed by using Stata software.Results 9RCTs involving 1 129subjects were included with 706subjects in DGMI treatment group and 423subjects in control group. (1) For acute ischemic stroke, DGMI group had superior effective rate compared to conventional therapy group (RR=1.19, 95％CI:1.09, 1.31, P＜0.000 1), improvement of neurologic impairments (SMD=3.23, 95％CI:2.87, 3.60, P＜0.000 1) and improvement of living quality (SMD=3.23, 95％CI:2.87, 3.60, P＜0.000 1). (2) For recovered stroke, DGMI group had better effective rate than Shuxuening injection group (RR=1.17, 95％CI:1.05, 1.30, P＜0.05) and improvement of neurologic impairments (SMD=-0.69, 95％CI:-0.88, -0.49, P＜0.000 1).There was no significant difference in adverse events between DGMI and control groups (P＞0.05).Conclusion DGMI had superior efficacy over control group for both acute ischemic stroke and recovered stroke.There was no significant difference in adverse events between these two groups.However, we still need better quality RCTs to confirm these results.

Objective To explore the layers of the facial and angular arteries between the orbital notch and the antegonial notch.Methods Anatomical studies were performed on 5 cases of adult cadavers (10 sides).Parallel lines were drawn on each cadaver through the position of the orbital incise,inner canthus,above of wing nose,nasal wing point,mouth angle,anterior incise.When the region of the arterial level changes,the parallel lines could be temporarily divided.The levels of the arteries that appeared on every horizontal line were recorded.Results The surface artery between the orbital notch (including the supraorbital notch) and the inner canthus (including the inner cantholes) was mainly in the orbicularis muscle layer.In the middle of the inner canthus and the highest point of the nasal wing,1/3 of the arteries were mainly walking in the orbicularis orbicularis muscular layer,and the next 1/3 were mainly located in the superficial layer or subcutaneous fat layer of the orbicularis oculi muscle.The highest point of the nose wing was found in the middle of the subcutaneous fat.The main line between the top of the nose and the lower part of the nose was deep in the subcutaneous fat.At the bottom of the nose,the main line was located in the fat layer of the zygomaticus,the zygomaticus maximus.In the middle and upper part of the nose and the middle of the mouth,a third of the main lines were walking in the lower fat layer (from shallow to deep) under the zygomaticus,and the next 1/3 segment was found in the subcutaneous fat deep.On the outside of the corner of the mouth,the main line was walking the subcutaneous fat layer or the lower fat layer of the platysma.Conclusions This study preliminarily clarifies the walking level of the facial artery from the upper orbital notch and the anterior notch of the angle.Bearing in mind the depth and the location of the vasculature within each zone,clinical practitioners can tailor their injection techniques to prevent vessel injury and avoid cannulation.

Fourteen neonatal diabetes mellitus(NDM)patients were recruited. 9 patients were treated with glyburide and the other 5 with insulin. ABCC8, KCNJ11, and INS genes were sequenced in 6 of them. Gene mutations were found in 2, 1, and 1 cases in these genes, respectively. One case with 6q24 hypomethylation and another without known mutation were also found. 8 out of 9 patients treated with glyburide reached euglycemia(88.9%). The other 5 patients with insulin therapy either died or lost contact. The results suggest that glyburide therapy is effective in neonatal diabetes mellitus, while insulin therapy may contribute to poor compliance.

OBJECTIVETo survey the current status of pubertal development of Chinese children and to compare the precocious puberty prevalence of different regions.

METHODSA cross-sectional epidemiological study was conducted on 18 707 children and adolescents aged 6≊18 y with male/female ratio of 9 812/8 895 from 6 representative geographical areas in China, including Beijing, Tianjin, Hangzhou, Shanghai, Chongqing and Nanning. The height, weight, waist circumference (WC), hip circumference(HC) and sexual maturation states (Tanner stages: breast stages for girls and testicular volume for boys) of children and adolescents were measured. Probit analysis was used to calculate the median age and 95% confidence interval (CI) for onset of breast and testicular development. The prevalence of precocious puberty of different regions and BMI, waist circumference of different groups were compared.

RESULTSBreast development before 8 y was observed in 2.91% of girls, and testicular volume 4 ml or more before 9 y was observed in 1.74% of boys. The median age of onset of Tanner stages 2 for breast development in girls was 9.69 y (95% CI: 9.63≊ 9.75); the median age of onset of puberty as indicated by Tanner stages 2 for testicular development in boys was 11.25 y (95%CI:11.19≊ 11.30). The prevalence of precocious puberty (43 girls and 37 boys) was 0.43% (80/18 707). The prevalence of precocious puberty in northern region was higher than that in southwest region (0.736% compared with 0.282% P<0.05). There was no difference in onset age of precocious puberty in girls among three regions; but the onset age of precocious puberty in boys was earlier in east China [(7.4±0.28)y]. The SD values of BMI and waist-to-hip ratio (W/H) in precocious puberty children were higher than those in the peer normal children. There was no difference in BMI,waist circumference and waist-to-hip ratio in the precocious puberty children among different regions.

CONCLUSIONThe current diagnostic criteria of precocious puberty are suitable for the children in the survey areas. The prevalence and the onset age of precocious puberty are various in different regions. A positive association between obesity and precocious puberty is found both in boys and girls.

Adolescent ; Age of Onset ; Child ; China ; epidemiology ; Cross-Sectional Studies ; Female ; Humans ; Male ; Obesity ; complications ; Prevalence ; Puberty, Precocious ; complications ; diagnosis ; epidemiology ; Sexual Development