Objective To observe the effect of Galectin -3 gene knock out to the mouse skin tumor growth and discuss the mechanism of inhibition of the mouse cutaneous tumor induced by 12 -o -tetrade-canoylphorbol-13-acetate which caused by Galectin -3 knock out.Methods The DMBA +TPA multi-step induced skin tumor in mice model were used to establish the skin cancer model .The control group was the same age wild type mice .We observed the inhibition of the mouse tumor growth by Galectin -3 knock out .In situ tumor cells were collected and cultured on soft agar for colony formation assay .The side population of the situ cancer cells was analyzed quantitatively by flow cytometry .Results 1.Compared with wild type mice group(group A), Galectin-3 knock out mice group ( group B) displayed a significant delay of the appearance of tumor .The tumor incidence and the average number of tumor per mice between group A and group B had obvious difference ( P<0.01).2.In vitro,data of soft agar colony formation assay showed that colony formation rate in group A are signif -icantly higher than group B(P<0.01).3.The collection and separation of A and B group in situ tumor cells ,u-sing flow cytometry instrument for in situ tumor cell side population quantitative analysis ,group A compared with group B had obvious difference(P<0.01).Conclusion The knock out of the Galectin -3 gene reduces the skin cancer stem cells ,inhibits tumor cell proliferation ,depresses chemical carcinogenesis of mice skin .Galectin-3 gene may become the new target for skin cancer chemotherapy .

Objective To explore the distribution and antimicrobial resistance of pathogens causing healthcare-associated infection(HAI) in a hospital,and provide theoretical basis for rational use of antimicrobial agents and control of HAL Methods HAI occurred in patients in this hospital between 2011 and 2015 were chosen,distribution of major pathogens causing HAI and antimicrobial resistance were analyzed.Results A total of 852 pathogenic strains were isolated,including 458 (53.75 ％) strains of gram-negative bacteria,259 (30.40 ％) fungi,and 135 (15.85 ％) gram-positive bacteria,the top 4 pathogens were Klebsiella pneumoniae (n =159,18.66 ％),Candida albicans (n =119,13.97％),Acinetobacter baumannii (n =81,9.51％),and Escherichia coli (n =74,8.69 ％).Isolation rate of gram-negative bacteria increased from 47.41 ％ in 2012 to 61.57％ in 2015,fungi increased from 24.78％ in 2011 to 34.45％ in 2014 (2015 was 27.95％),gram-positive bacteria decreased from 26.55％ in 2011 to 10.48％ in 2015.Resistance rates of Escherichia coli and Klebsiella pneumoniae to imipenem and meropenem were 8.11％-12.58％;Acinetobacter baumannii had the lowest resistance rate to cefoperazone/sulbactam(23.46％),resistance rate of Candida albicans to fluconazole was up to 59.66％,vancomycin-resistant gram-positive bacteria was not found.Conclusion The major pathogens causing HAI are opportunistic pathogens,antimicrobial resistance is serious,the species and distribution varies with different years,it is necessary to enhance specimen detection rate,strengthen the monitoring on pathogen resistance,and use antimicrobial agents rationally.

AIM: To study the effects of insulin on the proliferation and function of osteoblasts and the relationship between insulin post-receptor change in osteoblasts and osteoblastic cell growth.METHODS: The effects of different levels of insulin on osteoblasts were assessed by MTT colorimetry.Osteocalcin in medium was measured by RIM.IGF-1 mRNA expression levels were determined by RT-PCR.The concentrations of free IGF-1 protein in serum-free medium were measured by ELISA.In addition,the protein level and phosphorylated protein of P~(44/42)MAPK were determined by Western blotting analysis.RESULTS: Insulin enhanced the proliferation of osteoblasts,depending on its dose and exposure time.Insulin at concentration of 10~(-7) mol/L showed the strongest effect,and the action attained the plateau phase beyond 96 h.The best concentration that stimulated synthesis of osteocalcin by insulin was 10~(-7) mol/L.When the insulin concentration beyond 10~(-7) mol/L,the osteocalcin concentration was decreased.Exposure time had no effect on insulin-stimulated synthesis of osteocalcin of osteoblastic cells.When the concentration of insulin reaches 10~(-6) mol/L,the IGF-1 mRNA expression stimulated by insulin was also decreased.The concentrations of free IGF-1 protein in insulin-stimulated groups were all higher than that in control group(P0.05).Insulin acute stimulation rapidly induced the activity of tyrosine phosphorylation of P~(44/42)MAPK.The degree of tyrosine phosphorylation of P~(44/42)MAPK was increased step by step along with the increasing doses of insulin from 0 to 10~(-7) mol/L(P

Objective To investigate the clinical value of assessment of the left ventricular function in pa-tients with post-hepatitis b cirrhosis using Tei index and its related parameters measured by Tissue Dopple Imaging. Methods Ninety-seven subjects were enrolled,with 43 ones in the normal control group,and 54 ones in the post-hepatitis b cirrhosis group. According to the Child-Pugh grading standard,patients in the post-hepatitis b cirrhosis group were divided into grade Child A(24 cases),grade Child B(15 cases)and grade Child c(15 cases). Color ul-trasonograph was used to detect the left ventricle Tei indexes of patients in each group,and comparison was made in the differences of the parameters among all groups. Results (1)The Tei index,IRT/ET and ICT/ET of left ven-tricular in the liver cirrhosis group were higher than those in the normal/control group,with significant differences between the two groups(P < 0.05). The Tei index,IRT/ET and ICT/ET of the left ventricular increased gradually in patients of grade Child A,B,and C,and significant differences were observed in Tei index,IRT/ET(P < 0.05), but no significant diference in ICT/ET was found.(2)There were no significant difference in LVEF and Ea/Aa be-tween the cirrhosis group and the normal control group. LVEF and Ea/Aa in grade Child C were significantly lower than those in the normal control group(P < 0.05). Conclusion Tei index is superior to conventional echocardiog-raphy on assessment of the left ventricular function in patients with liver cirrhosis. Patients with higher the Child-Pugh score may be suffered from the worse the left ventricular function,especially the diastolic dysfunction.

Objective ToMeasureofleftventricularregionalsystolicfunctioninpatientswithcirrhosisby2DGSTItechnology.To exploretheclinicalvalueofleftventricularsystolicfunctionindifferentChildGPughgradedlivercirrhosispatientsanditscorrelation withthedegreeoflivercirrhosis.Methods Toselecting60casesoflivercirrhosispatientsincirrhosisgroup.AccordingtotheChildGPugh classification,thepatientsinA,BandCgradesweredividedintoA,BandCgroups.Toselecting30outpatientswithphysicalexaminations, whoseageandgendermatchedwithcirrhosispatientsatthesameperiodasnormal/controlgroup.Measuredtheconventionalindexes, andanalysedtwoGdimensionaldynamicimagesoftwoGchamber,threeGchamber,fourGchamberviewofapical.Thedifferencesinstrain indexandrelatedparametersbetweenthecirrhosisgroupsandthenormalcontrolgroupwerecompared.Results (1)Betweenliver cirrhosisgroupandcontrolgroup,therewerestatisticaldifferencesinheartrate,interventricularseptumthicknessandleftventricular enddiastolicvolume(P＜0.05).leftventricularejectionfraction wassignificantlylowerintheChildCgroupthanintheChild A groupandtheChildBgroup(P＜0.05),andwassignificantlyhigherintheChildBgroupthanthecontrolgroup(P＜0.05).Withthe increaseofChildGPughscoreinlivercirrhosisgroup,interventricularseptumthickness,leftatrialanteroposteriordiameter,leftventricular enddiastolicdiameterandleftventricularenddiastolicvolumeweregraduallyincreasing.Therewerestatisticallysignificantdifferences betweensomegroups(P＜0.05);(2)Longitudinalsystolicpeakstrainofeachsegmentofleftventricularwallwasgenerallylowerin thelivercirrhosisgroupthanthecontrolgroup,andthestrainin bothgroupsshowedincreasingtrendfromthebasaltotheapical segments.There werestatisticallysignificantdifferences between partialsegment(P＜0.05);(3)WiththeincreaseofChildGPugh scoreincirrhosisgroup,longitudinalsystolicpeakstrainofthe leftventricularwallgraduallydecreased.ChildCgroupcomparedwithothergroups,theleftventricularlongitudinalstrainaverage valuedifferencewasstatisticallysignificant(P＜0.05).Therewerestatisticallysignificantdifferencesinpartialsegmentbetweenthe othergroups(P＜0.05).Conclusion (1)2DGSTItechnologycanbesensitiveandreliabletodetectsubGclinicalmyocardialregional systolicdysfunctioninpatientswithcirrhosis;(2)LeftventricularsystolicfunctionisnegativelycorrelatedwithChildGPughgradein patientswithlivercirrhosis.

Objective K93T point mutation exists in the quinoid dihydropteridine reductase ( QDPR) of OLEFT rats which catalyzes QDPR into tetrahydrobinopterin(BH4), while dihydrofolate reductase(DHFR) can reduce QDPR to BH4, which implies crosstalk between hydrobiopterin and folate metabolism.By investigating the influence of QDPR expression on DHFR expression of NRK-52E cells, the article aimed to find out the possible underlying mechanism of QDPR gene in diabetic nephropathy ( DN). Methods Western blot was performed to identify the expression level in NRK-52E cell under high glucose ambience and DHFR pro-tein expression of OLETF rats.NRK-52E cells were transfected by the lentivirus to establish no-load overexpression, overexpressed QDPR and knockdown QDPR models.Each group was given 5.4 mmol/L normal sugar medium and 30mmol/L in high glucose ambi-ence for 72 hours'cell cultivation to simulate DN model.Observation was made on the influence of QDPR gene expression levels on DHFR in high glucose ambience. Results The western blot analysis revealed that DHFR protein decreased in NHG group( [0.33 ± 0.16] vs [0.64 ±0.5], P<0.05) and OLETF rats cortex ([0.56 ±0.16] vs [1.03 ±0.12], P<0.01).In high glucose ambi-ence, compared with LV-OCON-HG group, the protein expression of DHFR was significantly decreased in LV-QDPR-HG group ([0.12 ±0.09] vs [0.63 ±0.08], P<0.01).No difference was found in the comparison of DHFR expression levels between LV-SHQDPR-HG and LV-SHCON-HG group. Conclusion DHFR protein expression decreases in NRK-52E cells of high glucose and LOLETF rat model, which suggests that DHFR protein plays an important role in the development of DN.QDPR overexpression leads to the decreased expression of DHFR, which implies that overexpressed QDPR influences the occurrence and process of DN by down-regulating DHFR expression level.

_ Objective_ To study whether quinoid dihydropteridine reductase ( QDPR ) expression level change can affect oxidative stress of NRK-52E renal tubular cells in a high glucose environment. Methods The NRK-52E model of overexpression, knockdown QDPR gene and respective control were constructed by lentivirus. All groups were given 5. 4 mmol/L and 30 mmol/L glucose culture medium respectively to imitate normal and high glucose condition. The level of superoxide anion ( O-2 ) was detected by flow cytometer dihydroethidium method. The protein expression level of superoxide dismutase 1 (SOD1)was tested by Western blot. Results QDPR over-expression can decrease O-2(P<0. 01)and SOD1(P<0. 05)levels in high glucose condition;QDPR knockdown increases O-2(P<0. 01) and does not change SOD1. Conclusion Under high glucose condition, overexpression of QDPR gene decreases NRK-52E cell oxidative stress. Knockdown QDPR gene increases NRK-52E cell oxidative stress. QDPR gene may influence the development of diabetic nephropathy by oxidative stress.

BACKGROUND: It has been believed mesenchymal stem cells (MSCs) play a role in treatment through paracrine mechanism. Various side effects such as embolism, tumorigenesis and immunological reaction caused by direct injection of MSCs can be avoided by extracting MSC lysate. However, there is a larger difference in current collection methods and standards of MSC lysate. OBJECTIVE: To compare repeated freeze-thaw and ultrasonication for the collection of lysate of MSCs. METHODS: Adipose-derived mesenchymal stem cells (ADMSCs) were isolated from the abdominal subcutaneous fat of healthy individuals, and purified with adherence screening method, followed by in vitro amplification using fetal bovine serum medium. The common surface makers of these cells were tested by flow cytometry (1×109, 2×109, 4×109/L). Repeated freeze-thaw and ultrasonication were employed for cell cytoclasis at three different densities respectively in saline and double distilled water, and a comprehensive comparison was performed on cytoclasis rate and the content of protein in cell lysate between the two methods. RESULTS AND CONCLUSION: (1) ADMSCs obtained from in vitro isolated human adipose tissue grew in a swirl or radial pattern with a homogenous size and neat arrangement. CD44, CD90, CD105 and other commonly used surface markers were highly expressed. (2) The study for optimization of lysate collection revealed that the higher cell density implicated a longer time for cell wall disruption and cytoclasis, as well as significantly increased cytoclasis rate. (3) BCA protein assay showed that the highest content of protein was obtained in saline solvent using ultrasonication method. Comprehensive analysis on the results leads to a conclusion that ultrasonication method with saline as the solvent is the optimized method for extraction of ADMSCs lysate, and the cell concentration of less than 4×109/L is recommended.

Objective:To explore the value of Hong′s single-stitch duct to mucosa pancreaticoenterostomy(HSDMP) in laparoscopic pancreaticoduodenectomy(LPD).Methods:The perioperative clinical data of 300 patients undergoing LPD admitted to the Second Department of Hepatobiliary Surgery in the First Hospital of Jilin University from April 2015 to March 2019 were retrospectively analyzed. The patients were categorized into pancreatic fistula group( n=43) and non pancreatic fistula group( n=257). according to the presence or absence of postoperative pancreatic fistula. Among them, pancreaticojejunostomy was performed with HSDMP in 210 patients, and 90 patients underwent traditional pancreaticoenterostomy. Univariate and multivariate logistic regression were used to analyze the risk factors for the development of pancreatic fistula after LPD, and the difference on the time of pancreaticojejunostomy and the incidence of postoperative pancreatic fistula were compared between HSDMP and traditional pancreaticojejunostomy. Results:Univariate analysis showed that the differences on BMI, abdominal operation history, pancreatic texture, and pancreatic duct diameter were statistically significant compared with non-pancreatic fistula group (all P value <0.05). Multivariate logistic regression showed that BMI ( OR1.180, 95% CI1.047-1.338, P=0.008) and pancreatic texture( OR=0.375, 95% CI 0.135-0.861, P=0.036) were independent risk factors for pancreatic fistula after LPD. Compared with traditional pancreaticojejunostomy, HSDMP was associated with shorter anastomosis time and low incidence of postoperative pancreatic fistula, but there was no statistical significance between the two groups. Conclusions:The independent risk factors for pancreatic fistula are high BMI and soft pancreatic texture. Compared with traditional pancreaticojejunostomy, HSDMP does not prolong LPD time and increase the incidence of postoperative pancreatic fistula.

Objective:To analyze the research status and development trend of pediatrician competence in China and abroad, for reference in promoting the pediatrician competence research in the country.Methods:Literature review on pediatric resident′s competency was made using CNKI, WanFang and PubMed databases, ranging from their establishment up to December 25, 2019. A descriptive statistical analysis was made on the literatures′ publication year, publication area, research object and application area among others, using Excel 2019 and EndnoteX8.Results:A total of 199 Chinese documents and 262 English documents were finally retrieved. From 2009, the number of publications increased year by year, of which the Pediatrics has topped the rest, followed by Academic Medicine and Chinese Journal of Medical Education Research. The publishing institutions were mostly universities and affiliated hospitals, among which the Children′s Hospital of Chongqing Medical University topped the rest. The keyword of the highest frequency was education and the researches focus on the competency application and modelling. Conclusions:The pediatric healthcare system is paying greater attention on competence researches, but China has not yet established an appropriate pediatric resident competency model fitting local conditions. Meanwhile, the researches are plagued by such setbacks as excessive concentration of resources, simple methods and lack of empirical studies. It is recommended to broaden the explorations in this regard so as to identify pediatric resident competency development models fitting China′s conditions.