Objective To study the effect of acute lymphoblastic leukemia (ALL) children bone marrow mesenchymal stem cells (MSCs) on the resistance of K562cell atd mechanism in vitro.Method MSCs were obtained from AL children bone marrow after derivation, cultivation and identification.The coculture of MSCs and K562 and K562 suspension were established.Effects of MSCs on the growth of K562 cells were investigated in vivo.The two kinds of cells treated with different concentration of adriamycin (ADM) and the rate of apoptosis was evaluated by flow cytometry.Cell cycle was determined by flow cytometry.RT-PCR was used to detect Bcl-2 and Bax in K562 cells.Result Compared with the cell growth curve of K562 alone, the K562 cell co-cultured with MSCs grew slower and the exponential phase of growth was not obvious.The apoptosis index of the K562 cells co- clutured with MSCs was (9.19 ±0.53)% examined by flow cytometry, and that of the K562 cells alone was 4.00 ± 0.37% respectively( P ＜ 0.05 ).The percentage of cells at G0/G1 phase was (50.2 ± 2.26) % and that at S phase was (37.03 ± 3.50) % in the group of K562 alone, but those of the K562 cells co - cultured with MSCs were (80.95 ± 3.83) % and ( 17.40 ± 1.50)% respectively( P ＜0.05).The result of RT-PCR suggested expression of Bcl-2/Bax of the K562 cell co-cultured with MSCs was higher than K562 alone.Conclusion ALL children MSCs suppressed the growth of K562 cell in vitro.Adhesion made K562 depress sensitive to ADM.The mechanism was perhaps caused by adhesion with MSCs, K562 cell cycle was changed and related to Bcl-2 gene high level expression.

Objective To analyze the metastatic rule of common hepatic artery lymph node of thoracic esophageal squamous cell carcinoma,and to investigate the strategies of common hepatic artery lymph node dissection.Methods The clinical data of 682 patients with esophageal squamous cell carcinoma who were admitted to the Cancer Hospital of Fudan University from May 2005 to December 2010 were retrospectively analyzed.The locoregional lymph node metastasis of thoracic esophageal squamous cell carcinoma,relationship between metastatic rates of common hepatic artery lymph node and clinicopathological factors and the postoperative complications were analyzed.The enumeration data were analyzed using the chi-square test.Results A total of 18 277 lymph nodes were dissected (27 lymph nodes per patient).The lymph node metastatic rate was 55.87％ (381/682),and the metastatic lymph node ratio was 7.87％ (1438/18 277).Lymph nodes adjacent to the cardia of stomach,laryngeal nerve,lesser curvature of stomach,cervical esophagus,left gastric artery had a higher metastatic rate,while common hepatic artery lymph node had a lower metastatic rate.All the common hepatic artery lymph node metastasis was accompanied with locoregional metastasis.A total of 1480 common hepatic artery lymph nodes were dissected (2 common hepatic artery lymph nodes per patient).Twenty-four patients had common hepatic artery lymph node metastasis,with the metastatic rate of 3.52％ (24/682) and the lymph node ratio of 2.16％ (32/1480).The common hepatic artery lymph node metastatic rates of upper,middle and lower esophageal squamous cell carcinoma were 2.33％ (1/43),3.76％ (16/425) and 3.27％ (7/217),with no significant difference (x2 =0.295,P ＞ 0.05).The common hepatic artery lymph node metastatic rates of patients in T1,T2 and T3 stages were 2.35％ (2/85),5.46％ (10/183) and 2.90％ (12/414),with no significant difference (x2 =2.850,P ＞ 0.05).The common hepatic artery lymph node metastatic rates of patients with high,moderate and poor differentiated esophageal squamous cell carcinoma were 0(0/63),3.50％ (16/457) and 4.94％ (8/162),with no significant difference (x2=3.259,P ＞ 0.05).The common hepatic artery lymph node metastatic rates of patients with diameter of tumor under 3 cm,3-5 cm and above 5 cm were 2.59％ (6/232),3.02％ (11/364) and 8.14％ (7/86),with significant difference (x2 =6.267,P ＜ 0.05).The common hepatic artery lymph node metastatic rates of patients in N0,N1,N2,N3 stages were 0(0/301),2.53％ (5/198),5.65％ (7/124) and 20.34％ (12/56),with significant difference (x2 =62.368,P ＜ 0.05).The common hepatic artery lymph node metastatic rates of patients in stage Ⅰ,Ⅱ,Ⅲ and Ⅳ were 0(0/62),1.78％ (6/337),5.06％ (13/257) and 19.23％ (5/26),with significant difference (x2=25.959,P ＜0.05).Two hundred and twenty-eight patients had postoperative complications with the complication rate of 33.43％ (228/682).The incidence of anastomotic fistula was the highest,which was 11.58％(79/682).Conclusions The metastatic rates of common hepatic artery lymph node in thoracic esophageal squamous cell carcinoma is the lowest.For patients suffered from esophageal cancer in stage I or the tumor diameter under 5 cm,the dissection of common hepatic lymph node can be ommitted in surgery.

Objective Totally MIIE with per-oral placement of anvil has been reported elsewhere,but MIIE with manual pursestring and per-thoracic port placement of anvil has been seldomly reported.The feasibility of the latter technique was proved in this study.Methods Patients with mid-lower thoracic esophageal cancer were prospectively treated with totally MIIE at Shanghai Cancer Center of Fudan University from Feberay 28,2013 to August 31,2013.Laproscopic intracorporeal construction of the gastric conduit and needle catheter J-tube were performed in the first stage of MIIE.In the second stage a hand sewn pursestring was made with endostitch system and the anvil of EEA stapler was inserted via the tenth inter costal port prior to the intrathoracic anastamosis.Short-term clinicopathologic outcomes were collected.Results 39 cases were treated with totally MIIE,media age 61 years,ranged 48-69 years,10 females and 29 males.There was 1 conversion to open surgery.The median duration of operation was 245 minutes.The median intraoperative blood loss was 210 ml.All the patients were margin negative and staged from pT1N0M0 to pT3N2M0.The average lymph node yields were 16.5 per patient.The median postoperative hospital stay was 7 days.There was no mortality.Perioperative morbidity occurred in 4 patients (10％).2 patients were complicated with late stage gastric paralysis which began 2 or 3 days after oral feeding and both recovered in 1 month.1 patient was with minor anastamotic leakage which was endoscopically demonstrated on the 14th day postoperatively and the patient recovered in 1 month post leakage.1 patient was complicated with severe pneumonitus and ARDS; the ICU stay of that case was 19 days and the recovered patient was discharged 27 days postoperatively.Conclusion MIIE with regular EEA stapler and intrathoracic anastamosis is feasible in patients with thoracic esophageal cancer.Prospective randomized clinical trials could be conducted to compare the open procedure and totally MIIE with regular EEA stapler.

Objective To observe the efficacy ofvitrectomy combined with internal limiting membrane (ILM) peeling and scleral shortening for myopic foveoschisis (MF).Methods Prospective and non-randomized concurrent control study.A total of 35 MF patients (35 eyes) were enrolled in this study.The patients were divided into 2 groups according to surgery,including group A (18 eyes) and group B (17 eyes),all received vitrectomy combined with ILM peeling,but group A also received scleral shortening.The best corrective visual acuity (BCVA) examination using the Snellen vision chart was converted to the minimum resolution logarithm (logMAR).Ocular axis length (AL) was measured by Zeiss IOL-Master or A-scan ultrasound (Quantel Medical,France).The maximal value of retinal foveoschisis (MxFT) was measured by frequency-domain optical coherence tomography (Heidelberg,Germany).Multifocal electroretinogram (mfERG) responses were obtained with the RETIscan system (Roland Consult,Gemany).There was no statistically significant difference between the two groups (P＞0.05) in age (t=0.460),AL (t=1.520),diopter (t=0.020),logMAR BCVA (t=-2.280),MxFT (Z=-4.179) and b-wave ERG amplitude (Z=-0.198).The changes of BCVA,AL,MxFT and b wave amplitude were followed-up for 3-12 months.Results At the last follow-up,the height of MF was decreased in 18 eyes of group A,and MF was completely disappeared in 4 eyes.The logMAR BCVA (t=7.272,5.951),MxFT (Z=-3.724,-3.622) and b-wave ERG amplitude (Z=-3.223,-3.243) in both groups A and B were statistically improved (P=0.000,0.000,0.000,0.000,0.001,0.001) compared to pre-operational results.There was significant difference of logMAR BCVA (t=-2.280) and MxFT (Z=-4.179) between the two groups (P=0.029,0.000).But there was no significant difference in the amplitude of b-wave (Z=-0.198,P=0.843).The AL in group A was shortened after surgery,the difference was statistically significant (t=10.017,P=0.000).During the follow-up,there was no ocular hemorrhage,endophthalmitis and other complications.Conclusion PPV combined with ILM peeling and scleral shortening can shorten AL significantly for MF patients,and gain relative normal anatomical structure of the fovea,thus improve the vision.

Objective:To investigate the preventive efficacy of pirfenidone in esophageal stent-related restenosis and the related underlying mechanisms.Methods:Twenty-four rats underwent esophageal stent placement were included in this study. The rats were randomly assigned to three groups, with 8 rats in each group. The three groups were set to receive placebo, 150 mg/kg pirfenidone and 300 mg/kg pirfenidone daily by oral administration for 28 days, respectively. Twenty-eight days after stent placement, the stented esophagi were harvested for histological examinations. The number of epithelial layers, the thickness of submucosal fibrosis, the percentage of granulation tissue area, the degree of inflammatory cell infiltration, the degree of collagen deposition, and the α-SMA staining scores were evaluated. One-way ANOVA was performed for the statistical comparison of the number of epithelial layers, the degree of inflammatory cell infiltration, the degree of collagen deposition and the α-SMA staining scores among these three groups. The Kruskal-Wallis H test was used for comparison of the thickness of submucosal fibrosis and the percentage of granulation tissue area among the three groups. Results:Gross pathological findings showed that both pirfenidone groups had significantly less luminal fibrotic tissue formation and restenosis than placebo group. The percentage of granulation tissue areas in placebo group, 150 mg/kg and 300 mg/kg pirfenidone groups were 57.23%±25.68%, 21.80%±6.65% and 12.18%±6.37%, respectively. Both pirfenidone groups showed significantly less granulation tissue areas than placebo group ( P<0.01). The degree of inflammatory cell infiltration, the degree of collagen deposition and the α-SMA staining scores were 3.28±0.55, 3.38±0.63 and 2.75±0.38 in placebo group, 2.30±0.46, 2.36±0.58 and 2.00±0.42 in 150 mg/kg pirfenidone group, and 1.86±0.38, 1.91±0.41 and 1.57±0.28 in 300 mg/kg pirfenidone group, respectively. Both pirfenidone groups showed significantly less inflammatory cell infiltration, collagen deposition and α-SMA staining scores than placebo group ( P<0.01). Conclusion:Pirfenidone can suppress esophageal stent-related restenosis in rats by significantly inhibiting inflammation, myofibroblast activation and proliferation, and fibrotic tissue formation.

OBJECTIVETo study the effect of DNase I on biofilm formation of Staphylococcus aureus.

METHODSThe growth curve of S. aureus was detected using a spectrophotometer. The adhesion of S. aureus was analyzed using flat colony counting method, and the biofilm formation was assayed using the 96-well crystal violet staining method.

RESULTSExposure to different concentrations of DNase I did not obviously affect the growth of S. aureus but significantly inhibit the formation of bacterial biofilms in a dose-dependent manner. DNase I inhibited the adhesion of S. aureus at different growth stages. When combined with antibiotics, DNase I resulted in a signi?cant decrease in the established bio?lm biomass compared to antibiotics or DNase I used alone.

CONCLUSIONDNase I can effectively inhibit biofilm formation of S. aureus and enhance the inhibitory effect of antibiotics against S. aureus biofilms.

OBJECTIVE: To observe the expression of EphA2, and investigate its correlation with the development, progression, invasion, metastasis, angiogenesis and prognosis in nasopharyngeal carcinoma(NPC). METHOD: Immunohistochemical staining was used to determine the expression level of EphA2 protein in 61 cases NPC and 20 cases chronic nasopharyngitis samples. The clinically pathological data and results of follow-up were collected. Microvessel density (MVD) was also measured by immunohistochemical staining method with CD34 in NPC. RESULT: The positive rate of EphA2 protein staining in NPC was 60.66% (37/61), while that in nasopharyngitis samples was 10.0% (2/20). The positive rates of EphA2 protein in NPC were 27.27% (3/11) in stage I, 56.25% (9/16) in stage II, 68.19% (15/22) in stage III, and 83.33% (10/12) in stage IV. The positive expressions of EphA2 in T1 + T2 and T3 + T4 with neck lymph node and distant metastasis were 58.33% (7/12) and 88.89% (16/18) respectively, while those in T1 +T2 and T3 + T4 without metastasis were 31.25% (5/16) and 50.00% (6/12) respectively. The cumulative survival of patients in the EphA2 positive group at 5 years was only 0.324 (12/37), while 0.500 (12/24) in the EphA2 negative group. The positive expression of EphA2 protein was correlated with the clinical stage, the neck lymph node metastasis and distant metastasis, and prognosis of NPC, respectively (P < 0.05). MVD in EphA2 protein positive group (45.32 +/- 4.91) was significantly higher than that in EphA2 protein negative group (28.69 +/- 3.99, P < 0.01). CONCLUSION EphA2 may play an important role in the development and progression of NPC. It is closely associated with the invasion, metastasis, angiogenesis and prognosis of NPC.
Adolescent ; Adult ; Aged ; Carcinoma ; Female ; Humans ; Lymphatic Metastasis ; Male ; Microvessels ; Middle Aged ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; blood supply ; metabolism ; pathology ; Neoplasm Staging ; Neovascularization, Pathologic ; pathology ; Prognosis ; Receptor, EphA2 ; metabolism ; Young Adult

Objective:To investigate the effect of long-chain non coding RNA (lncrna) loc730101 in the proliferation, invasion and migration of U2OS cells, and its mechanism.Methods:From February, 2019 to October, 2019, U2OS cells cultured in vitro were divided into control group (normal culture), negative group (transfection nega- tive control), and interference group (transfection of interference sequences targeting LOC730101). The expression of LOC730101 in cells was detected by reverse transcription-polymerase chain reaction (RT-PCR). Cell proliferation ac tivity was tested by methylthiazolyldiphenyl-tetrazolium bromide (MTT) method. Cell clone formation rate was mearused by plate clone formation test. Cell cycle distribution was tested by flow cytometry. Cell invasion and migra- tion were examed by Transwell chamber. The expression levels of epithelial-mesenchymal transition-related proteins Vimentin, N-cadherin, E-cadherin, and Wnt/β-catenin signaling pathway related proteins in cells β-catenin, c-Myc, cyclin D1(CyclinD1) and matrix metalloproteinase-7(MMP-7) proteins were detected by Western blotting method. The date was statistical analysed and considered as statistically significant when P<0.05. Results:Compared with the control group, the expression level of LOC730101 (0.25±0.03 and 1.00±0.06) in interference group and control group, respectively. The same below), cell survival rate [(57.65±3.26)% and (100.00±7.39)%], clone formation rate [(13.03± 2.12)% and (25.35±3.58)%], number of invasive cells(51.36±3.48 and 92.85±6.62), number of migrating cells (77.15± 5.05 and 136.92±15.35), the percentage of cells in S phase [(20.54±2.15)% and (28.15±2.38)%] and G 2/M phase [(16.87±2.12)% and (23.36±3.12)%], as well as the expression level of Vimentin (0.52±0.04 and 1.17±0.13), N-cadherin (0.31±0.03 and 0.65±0.04), β-catenin (0.42±0.03 and 0.73±0.04), c-Myc (0.29±0.03 and 0.65±0.03), CyclinD1 (0.26± 0.02 and 0.58±0.04), MMP-7 protein (0.55±0.03 and 0.86±0.06) was decreased significantly ( P<0.05), while the per- centage of cells in G 0/G 1 phase [(62.62±5.15)% and (48.46±3.65)%] and the expression level of E-cadherin protein(0.82± 0.06 and 0.38±0.03) were increased significantly in the interference group ( P<0.05). But there was no significant differ- ence in each index in the negative group ( P>0.05). Conclusion:Reduce the regulation of LOC730101 can inhibit the proliferation, invasion and migration of U2OS cells, and its mechanism may be related to the inhibition of Wnt/β-catenin signaling pathway.

Objective: To present the evolutionary path of Chinese Medical Ethics, analyze the development vein of periodical, explore the journal' s research focuses, and provide reference for related personnel to understand the development of the current research status in the field through visualization technology. Methods:Using biblio-metric method,taking "China National Knowledge Infrastructure" as source of data collection, we used the visual-ization software CiteSpace to draw scientific knowledge maps and analyzed literatures published from 1990 to 2014 in Chinese Medical Ethics. Results:The annualvolume of journal articles fluctuated upwards,with peaks in 1992, 2000 and 2009, and of which the most was in 2009, with 393 articles. "Medical Ethics" and "Doctor-patient Relationship" were hotwords in this field. The publications of domestic and foreign scholars promoted the interna-tional exchange and the development of Chinese medical ethics, and the most productive institutions were often col-leges or universities. Conclusion:Chinese Medical Ethics has effectively promoted the development of bioethics in China, more and more scholars are involved in the relative research of medical ethics, and the old, middle and young scientists and research teams inherit, cooperate and develop with each other. The cross-regional, inter-a-gency and interdisciplinary collaboration remains very limited, whichwill become the important factorinfluencingthe development of the field of Chinese medical ethics in the future.

Objective:To evaluate the feasibility and safety of a novel transjugular intrahepatic portosystemic shunt (TIPS) puncture set for transjugular intrahepatic portal puncture in swine.Methods:Thirteen domestic swine were randomly divided into experimental group ( n=7) and control group ( n=6) and received transjugular intrahepatic portal puncture with the novel puncture set and the R?sch-Uchida Transjugular Liver Access Set (RUPS-100), respectively. Three swines in each group were randomly selected and sacrificed immediately after the procedure and the rest were sacrificed 1 week later. The intraoperative technical success rate, puncture attempts, procedure time, fluoroscopy time, vital signs and occurrence of complications related to the procedure as well as the changes of liver and kidney function before, immediately after and 1 week after the procedure were compared between two groups. Independent sample t test, paired t test, Mann-Whitney U test, chi-square test or Fisher′s exact test were performed to compare the differences between groups. Results:The technical success rate was 100% for both groups. No significant difference in portal puncture attempts [2.0 (1.0, 5.0) vs. 2.0(1.0, 5.5), P>0.999], procedure time [(47.7±20.6) vs. (52.5±28.0)min, P=0.729] and fluoroscopy time [(725.1±489.2) vs. (763.7±562.4)s, P=0.897] was observed between two groups. In addition, no significant difference of the major liver and kidney function between two groups before, immediately after and 1 week after procedure ( P>0.05 at all time points). No significant difference of the major liver and kidney function change in two groups during perioperative period was identified (all P>0.05). Furthermore, no significant difference of non-target puncture occurrence was observed between two groups ( P>0.999). In the meantime, no significant difference of occurrence of small hematomas around hepatic hilar was discovered among swine sacrificed immediately after procedure between two groups ( P>0.999). No other major complications were identified in either group. Conclusion:The novel TIPS puncture set is feasible and safe to perform transjugular intrahepatic portal puncture in swine.