Research advances in molecular targeted therapy for gastroenteropancreatic neuroendocrine neoplasms / 中华消化外科杂志

Neuroendocrine neoplasms, commonly deriving from neuroendocrine cells of gastrointestinal tract and pancreas, are remarkably heterogeneous. As for gastroentero-pancreatic neuroendocrine neoplasms, surgery is the first choice for treatment, whereas molecular targeted therapy provides a new treatment option for patients with local advancement or metastases. Drugs for molecular targeted therapy used clinically include somatostatin analogs, mammalian target of rapamyoin pathway inhibitors, tyrosinekinase inhibitors, immunotherapy and so on. Research findings from experiments and clinic trials will possibly provide new therapeutic methods for molecular targeted therapy in the future.

Associations between Tensin1 protein and clinicopathological characteristics and prognoses of gastric cancer patients / 中华消化外科杂志

Objective To investigate the associations between expression of Tensin1 protein and clinicpathological characteristics and prognoses of gastric cancer (GC) patients.Methods The retrospective casecontrol study was conducted.The clinicopathological data of 163 GC patients who were admitted to the Affiliated Hospital of Jiangsu University between July 31,2011 and December 31,2013 were collected.The GC tissues and adjacent normal tissues were taken to paraffin imbedding,and then were detected by immunohistochemistry.Observation indicator:(1) expressions of Tensinl protein in GC tissues and adjacent normal tissues;(2) association between expression of Tensinl protein in GC tissues and clinicopathological characteristics;(3) followup and survival situations;(4) prognostic factors analysis.Follow-up using telephone interview was performed to detect survival up to January 1,2017.Measurement data with skewed distribution were described as M (range).Count data were analyzed using the chi-square test or pairing chi-square test.The survival curve and rate were respectively drawn and calculated using the Kaplan-Meier method,and Log-rank test was used for survival analysis.The univariate analysis and multivariate analysis were done using the COX roportional hazard model.Results (1) Expressions of Tensinl protein in GC tissues and adjacent normal tissues:immunohistochemistry showed that Tensinl protein in GC tissues and adjacent normal tissues mainly expressed in cytoplasm.Of 163 patients,154 (66 with high expression and 88 with low expression) and 9 had respectively positive and negative expressions of Tensinl protein in GC tissues;79 (37 with high expression and 42 with low expression) and 84 had respectively positive and negative expressions of Tensinl protein in adjacent normal tissues,with statistically significant differences in positive expression ratio and expression levels (x2=64.65,12.93,P＜0.05).(2) Association between expression of Tensinl protein in GC tissues and clinicopathological characteristics:high expression rate of Tensinl protein in GC tissues were respectively 31.34％ (21/67) in GC patients with tumor metastases and 46.88％ (45/96) in GC patients without tumor metastasis,with a statistically significant difference (x2 =3.95,P＜0.05).(3) Follow-up and survival situations:all the 163 patients were followed up for 3.3-64.7 months,with a median time of 28.7 months.The 3-year cumulative disease-free survival rate and cumulative overall survival rate in GC tissues were 63.12％,74.22％ in 66 patients with high expression of Tensinl protein and 47.30％,55.74％ in 97 patients with low and negative expressions of Tensin1 protein,showing statistically significant differences in above indicators (x2 =4.58,4.11,P＜0.05).Survival analysis of subgroups showed that 3-year cumulative disease-free survival rate in GC tissues of patients with maximum tumor dimension ≥ 5 cm,nerve and / or vascular invasions and stage Ⅲ of TNM staging were 45.98％,62.79％,52.75％ in patients with high expression of Tensin1 protein and 18.11％,31.10％,32.80％ in patients with low and negative expressions of Tensin1 protein,with a statistically significant difference (x2 =5.85,7.89,4.96,P＜0.05).The 3-year cumulative overall survival rate was respectively 66.00％,75.75％,67.93％ in patients with high expression of Tensin1 protein and 30.74％,40.15％,44.67％ in patients with low and negative expressions of Tensinl protein,with statistically significant differences (x2 =7.59,9.62,4.32,P ＜ 0.05).(4) Prognostic factors analysis:results of univariate analysis showed that maximum tumor dimension,histological grade,nerve and / or vascular invasions,postoperative TNM staging,postoperative adjuvant chemotherapy and expression of Tensin1 protein were related factors affecting prognoses of GC patients [hazard ratio (HR) =3.66,2.45,2.17,3.36,0.41,0.54,95％ confidence interval (CI):2.09-6.41,1.43-4.19,1.17-4.04,1.52-7.41,0.23-0.72,0.31-0.96,P＜0.05].Results of multivariate analysis showed that maximum tumor dimension ≥ 5 cm and grade Ⅲ of histological grade were independent risk factors affecting prognoses of GC patients (HR=3.21,2.17,95％CI:1.63-6.32,1.18-3.99,P＜0.05),and postoperative adjuvant chemotherapy and high expression of Tensin1 protein were independent protective factors affecting prognoses of GC patients (HR=0.50,0.44,95％CI:0.28-0.90,0.24-0.82,P＜0.05).Conclusion High expression of Tensin1 protein may inhibit GC metastasis,and it is also an independent protective factor affecting prognoses of GC patients.

Prognostic analysis and clinicopathological features of 20 patients with appendiceal neuroendocrine neoplasms / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate clinicopathological features and prognostic factors of appendiceal neuroendocrine neoplasms(a-NEN).</p><p><b>METHODS</b>Clinical data of 20 patients diagnosed with a-NEN at Zhongshan Hospital of Fudan University between January 2000 and December 2016 were retrospectively analyzed. Pathological diagnosis was based on the WHO classification criteria of digestive system tumors (2010 edition). Based on the mitotic count and Ki-67 index, a-NENs were divided into grade 1 neuroendocrine tumor (NET G1), grade 2(G2) NET G2) and grade 3 (neuroendocrine carcinoma, NEC). Some special types of a-NEN (e.g. goblet cell carcinoid) and mixed adenoneuroendocrine neoplasms were classified as mixed adenoneuroendocrine carcinoma (MANEC). Follow-up was conducted by telephone or return visits. Univariate analysis was carried out using the Kaplan-Meier method, and the log-rank test was used to draw survival curves.</p><p><b>RESULTS</b>Of 20 patients, 14 were male and 6 were female with median age of 54 years. Seventeen cases presented acute right lower quadrant abdominal pain, 1 chronic right lower quadrant abdominal pain, 1 persistent abdominal discomfort with outburst whole abdominal pain and 1 was found during body check without symptoms. Twenty cases comprised 8 G1 patients, 4 G2 patients, 3 G3 patients, and 5 MANEC patients. When diagnosed, there was 1 patient with liver metastasis, 1 patient with abdominal and pelvic metastases, and 2 patients with postoperative pathological findings of lymph node metastasis. Six patients underwent appendectomy, 12 underwent right hemicolectomy, 1 underwent right hemicolectomy plus small intestine resection, and 1 underwent partial hepatectomy plus right hemicolectomy. The follow-up time was 7-187 months(average, 36 months). The total 1- and 3-year survival rates were 94.7% and 60.2%, respectively. Univariate analysis showed that age >50 years (χ=7.036, P=0.008), pathology grade as MANEC (χ=5.297, P=0.021), and metastasis (χ=6.558, P=0.010) indicated lower 5-year survival rate.</p><p><b>CONCLUSIONS</b>Most a-NEN patients have no typical symptoms, and the main complaint at consultation is acute right lower quadrant abdominal pain. Prognosis is poor for patients with age >50 years, MANEC pathology grade and metastasis.</p>

Clinical effect of mirabilite external application and neostigmine injection at Zusanli acupoint combined with Xuebijing intravenous injection on patients with severe sepsis / 中国中西医结合急救杂志

ObjectiveTo observe the clinical effects of mirabilite external application and neostigmine injection at Zusanli acupoint combined with Xuebijing intravenous injection on patients with severe sepsis.Methods A history-prospective controlled study was conducted. Patients with severe sepsis admitted to the Fourth People's Hospital of Jiangsu University from January 2012 to November 2015 were enrolled. Twenty-one cases admitted from January 2014 to November 2015 were assigned as a research group and treated with application of mirabilite external application, Zusanli acupoint injection of neostigmine combined with intravenous Xuebijing injection; 22 patients with Xubijing treatment from January 2012 to November 2015 were included in a Xubijing group; 21 patients with routine therapy from January 2012 to November 2013 were included in a control group. The changes of white blood cell count (WBC), C-reactive protein (CRP), procalcitonin (PCT), mean arterial pressure (MAP), oxygenation index (OI), serum creatinine (SCr), acute physiology and chronic health evaluation Ⅱ(APACHEⅡ) score, sequential organ failure assessment (SOFA), gastrointestinal function score, blood platelet count (PLT), plasma prothrombin time (PT), activated partial thromboplastin time (APTT), and D-dimmer in three groups before and after treatment were observed; the length of stay in ICU and the 28-day mortality were compared among the three groups. Results Compared with those before treatment, WBC, PCT, CRP, SCr, APACHE Ⅱ score, SOFA score, gastrointestinal function score, APTT, PT, and D-dimer were all obviously lower after treatment for 7 days in various groups; OI, MAP, PLT were significantly higher, and the improvement degree of WBC, PCT, CRP, SCr, APACHE Ⅱ score, SOFA score, gastrointestinal function score, OI in research group was more significant than those of control group and Xubijing group [WBC (×109/L): 7.52±0.75 vs. 12.87±4.13, 10.88±0.66, PCT (μg/L): 1.14±0.55 vs. 6.32±1.39, 3.47±1.94, CRP (mg/L): 21.0±9.2 vs. 65.0±13.6, 35.0±13.9, OI (mmHg, 1 mmHg = 0.133 kPa): 357.0±20.4 vs. 295.0±20.4, 309.0±21.4, SCr (μmol/L): 7.89±2.35 vs. 14.33±9.17, 11.27±4.65, APACHE Ⅱ score: 10.38±0.75 vs. 18.27±2.57, 13.09±4.10, SOFA score: 1.05±0.66 vs. 6.01±2.33, 3.26±1.03, gastrointestinal function score: 0.31±0.11 vs. 2.01±0.46, 1.85±0.29, all P 0.05]. Conclusions The treatment of mirabilite external application and neostigmine injection at Zusanli acupoint combined with Xuebijing intravenous injection can obviously improve the clinical symptoms, blood coagulation indexes and organ functions, reduce the levels of inflammatory indexes, shorten the time of the length of stay in ICU and elevate the survival rate of patients with severe sepsis.

Influence of RNA interference in p53 gene on the expressions of genes involved in ultraviolet B-induced premature senescence and photocarcinogenesis in human skin fibroblasts / 中华皮肤科杂志

Objective To evaluate the effect of RNA interference in p53 gene on the expressions of genes involved in ultraviolet B (UVB)-induced premature senescence and photocarcinogenesis in human skin fibroblasts (HSFs).Methods A previously established HSF cell clone with repressed expression of p53,which was named as HSF-p53,was cultured and irradiated with a subcytotoxic dose (10 mJ/cm2) of UVB once a day for five consecutive days.The HSFs with normal expression of p53 served as the control.Subsequently,β-galactosidase (SA-β-gal)-staining was performed to estimate the degree of senescence,quantitative real-time PCR array was performed to determine the mRNA expressions of photocarcinogenesis-and senescence-associated genes,including p53,p21,p19,p16,pRb,fibronectin,osteonectin,smooth muscle 22 (SM22),bax,bcl-2,hypoxia-inducible factor-1 α (HIF-1α),vascular endothelial growth factor(VEGF),and human double minute-2 (hdm2).Statistical analysis was carried out by Student's t test using the software SPSS 10.0.Results The percentage of SA-β-gal-positive cells in irradiated HSF-p53 was 19.70％ ± 0.85％,significantly higher than that in unirradiated HSF-p53 (12.77％ ± 0.81％,t =6.45,P ＜ 0.05),but lower than that in irradiated control HSFs (50.48％ ± 5.30％,t =7.86,P ＜ 0.05),and similar to that in unirradiated control HSFs (18.50％ ± 0.45％,t =2.57,P ＞ 0.05).Compared with the control HSFs,the HSF-p53 showed decreased expressions of p21,p19,fibronectin,osteonectin,SM22 and bax genes (all P ＜ 0.05),but increased expressions of bcl-2,HIF-1α,VEGF and hdm2 genes (all P ＜ 0.05),and a similar expression of p16 gene (P ＞ 0.05); the repeated UVB radiation significantly promoted the expressions of p16 and pRb genes (both P ＜ 0.05),but had no obvious effect on the expressions of the other genes in HSF-p53 compared with unirradiated HSF-p53 (all P ＞ 0.05).Conclusions The inhibition of p53 expression may decelerate the UVB-induced premature senescence in HSFs,which may be involved in the p53-dependent tumor suppression.

Analysis of an acellular pigskin based nerve scaffold / 生物工程学报

A scaffold fabricated with lysine/nerve growth factor (NGF)/poly (lactic acid coglycolic acid) copolymer (PLGA) and acellular pigskin was evaluated in vitro as a potential artificial nerve scaffold. Properties of the scaffold such as microstructure, mechanical property, degradation behavior in PBS and water, Schwann cell adhesion property, and release of NGF were investigated. Results showed PLGA had permeated into the porous structure of acellular pigskin; its breaking strength was 8.308 MPa, breaking extensibility was 38.98%, elastic modulus was 97.27 MPa. The porosities of the scaffold ranged from 68.3% to 81.2% with densities from 0.62 g/cm3 to 0.68 g/cm3. At 4 weeks of degradation in vitro, maximum mass loss ratio was 43.3%. The release of NGF could still be detected on the 30th day, and its accumulative release rate was 38%. Lysine added into the scaffold neutralized the acidoid preventing degradation of PLGA to maintain a solution pH value. Schwann cells had grown across the scaffold after co-cultivation for 15 days. These in vitro properties of the pigskin based composite might indicate its potentiality to be an artificial nerve scaffold.

Influence of RNA interference-induced repression of p53 expression on senescence in human skin fibroblasts / 中华皮肤科杂志

Objective To establish a cell line with repressed expression of p53 by transfecting a plasmid construct expressing short hairpin RNA(shRNA)targeting p53 into human skin fibroblasts(HSFs),and to evaluate the effect of repression of p53 expression on the senescence in HSFs.Methods The eukaryotic expressing plasmid pGCsi-p53 containing shRNA targeting p53 gene was transfected into HSFs with lipofectamine.Subsequently,the cells were selected by G418,and resistant cell clones were chosen and expanded.Reverse transcription-PCR and real time fluorescence-based quanitative PCR were performed to determine the expression of p53 gene,and Western blot to detect the expression of p53 protein in HSFs.The senescence in HSFs was evaluated by SA β-gal staining,and cell proliferation by methyl thiazolyl tetrazolium(MTT)assay.Results A HSF clone with repressed expression of p53 was established successfully.The expressions of p53 mRNA and protein were downregulated in transfected HSFs compared with untransfected HSFs(0.09 ± 0.03 vs.0.32 ± 0.04,0.11 ± 0.04 vs.0.84 ± 0.05,both P ＜ 0.01).The percentage of senescent cells was 13.47％ ± 1.01％ in the transfected HSFs,significantly lower than that in untransfected HSFs(18.10％ ± 0.66％,P ＜ 0.05).As MTT assay showed,the proliferation was accelerated in transfected HSFs compared with untransfected HSFs(P ＜ 0.05).Conclusions The repression of p53 expression decelerates the senescence in HSFs,but promotes the proliferation of HSFs.

Influences of extracellular matrices secreted by ultraviolet B-induced senescent fibroblasts on ERK signaling in HaCaT cells / 中华皮肤科杂志

Objective To explore the influences of extracellular matrices (ECM) secreted by ultraviolet B (UVB)-induced senescent fibroblasts on the proliferation of and extracellular signal-regulated kinase (ERK) signaling in HaCaT cells. Methods Fibroblasts were irradiated with UVB of 15 mJ/cm2 once daily for 5 days to induce premature senescence, which was identified by SA-β-gal staining 72 hours after the last irradiation.HaCaT cells were divided into 3 groups and inoculated into plates coated with extracellular matrices secreted by non-senescent (PRE-ECM) or senescent fibroblasts (SIPS-ECM) or into uncoated plates (NON-ECM), fol-lowed by additional culture. U0126, an inhibitor of ERK1/2, was used to treat the HaCaT cells 1 hour before inoculation. Then, MTT assay was carried out to detect the proliferation of HaCaT cells after a 3-day culture,Western blot to assess the phosphorylation of ERK at 0.5, 1, 2 and 4 hours after the inoculation, flow cytometry to analyse cell cycle and apoptosis after 24 hours of culture. Results The most rapid and intense phosphory-lation of ERK was observed in SIPS-ECM group. Inhibiting the activation of ERK pathway with U0126 could completely suppress the promoting effect of ECM from senescent fibroblasts on the proliferation of HaCaT cells.After the blocking of ERK activation, the proportion of HaCaT cells in S and G2/M phase decreased from 37.40%, 41.34% and 43.31% to 29.41%, 36.48% and 39.96%, respectively, in NON-ECM, PRE-ECM and SCIP-ECM group. Conclusion The ECM produced by UVB-induced senescent fibroblasts promote the prolifera-tion of HaCaT cells via inducing the phosphorylation of ERK.

Study on the Superantigen Production by Skin-Colonizing Staphylococcus aureus in Patients with Atopic Dermatitis and Eczema / 中华皮肤科杂志

Objective To determine the potential impact of superantigens produced by skin-colonizing Staphyiococcus aureus in patients with atopic dermatitis and eczema. Methods Of 117 patients with atopic dermatitis and 199 with eczema, 140 Staphyiococcus aureus strains were isolated from the skin specimens. Superantigens were detected with reverse passive latex agglutination. Results Among 140 Staphyiococcus aureus strains, 60 (42.9%) produced superantigens, among which 43 produced one kind of superantigens only and 17 produced at least two kinds. Of strains isolated from atopic dermatitis, 51.5% produced superantigens and no significant difference was seen in superantigen production between lesional and non-lesional strains in atopic dermatitis. Of strains isolated from eczema patients, 34.7% (all were lesional strains) produced superantigens. The positive rates of total superantigens, lesional superantigens and toxic shock syndrome toxin-1 production were all higher in the strains from atopic dermatitis than in those from eczema. Conclusions Superantigen production by skin-colonizing Staphyiococcus aureus probably plays a more important role in atopic dermatitis than that in eczema. However, further studies are necessary to validate its importance.