OBJECTIVE To investigate the in vitro inhibitory mechanism of berberine on the proliferation of tumor stem cells and evaluate its in vivo safety. METHODS Flow cytometry was used to select tumor stem cells from mouse skin melanoma B16F10 cells； CD44， CD133， Nanog homologous box protein （NANOG） and octamer-binding transcription factor 4 （OCT4） were used as indicators to characterize tumor stem cells. Tumor stem cells were divided into control group， all-trans retinoic acid （ATRA） group， and berberine group， and the CCK-8 method was used to detect the effects of berberine on the viability of tumor stem cells； flow cytometry was adopted to detect cell apoptotic rate， the proportion of CD44+/CD133+ and the positive cell rate of sex determining region Y box protein 2 （SOX2）； the morphological changes of tumor balls were recorded after treatment with berberine； the morphology of cell pyroptosis in each group was recorded， and the release rate of lactate dehydrogenase （LDH） was detected； Western blot assay was adopted to detect the expressions of pyroptosis-related protein gasdermin E （GSDME）， GSDME- N， caspase-3 and cleaved caspase-3. Preliminary evaluation of in vivo safety of berberine was conducted by using zebrafish embryo toxicity experiments. RESULTS Compared with B16F10 cells， the proportion of CD44+/CD133+ cells in tumor stem cells and the fluorescence intensity of NANOG and OCT4 were significantly increased （P＜0.000 1）. The half-inhibitory concentration of berberine to tumor stem cells was 50.98 μmol/L. Compared with the control group， the apoptotic rate of cells in the berberine group was significantly increased， while the proportion of CD44+/CD133+ cells and the rate of SOX2 positive cells were reduced significantly （P＜0.000 1）； tumor stem cell spheroids were atrophied， with partial cell death. After treatment with berberine， tumor stem cells exhibited swelling in their outermost layer， the release rate of LDH of cells was significantly increased and the release rate of LDH increased with increasing dose； the protein expressions of GSDME-N and cleaved-caspase-3 of cells in berberine 20， 40 μmol/L groups were significantly increased， and the protein expressions of GSDME and caspase-3 were significantly reduced （except for berberine 20 μmol/L group， P＜0.05）. The embryonic development of zebrafish treated with berberine was almost unaffected， and the survival rate of embryo reached 100%， with no obvious abnormalities observed. CONCLUSIONS Berberine has good activity against the proliferation of tumor stem cells， and its mechanism of action may be related to activating GSDME and promoting cell pyroptosis； berberine has good in vivo safety.

RNAs are involved in the crucial processes of disease progression and have emerged as powerful therapeutic targets and diagnostic biomarkers. However, efficient delivery of therapeutic RNA to the targeted location and precise detection of RNA markers remains challenging. Recently, more and more attention has been paid to applying nucleic acid nanoassemblies in diagnosing and treating. Due to the flexibility and deformability of nucleic acids, the nanoassemblies could be fabricated with different shapes and structures. With hybridization, nucleic acid nanoassemblies, including DNA and RNA nanostructures, can be applied to enhance RNA therapeutics and diagnosis. This review briefly introduces the construction and properties of different nucleic acid nanoassemblies and their applications for RNA therapy and diagnosis and makes further prospects for their development.

Objective:To investigate the value of total laparoscopic simultaneous resection for left-sided colorectal cancer (CRC) and synchronous liver metastases (SLM).Methods:A retrospective analysis of the clinical data of patients with left-sided CRC and SLM who underwent simultaneous resection in the Shanghai Cancer Center, Fudan University from March 2014 to December 2017. The patients were divided into laparoscopy group, open surgery group and hybrid surgery group. The intraoperative information, postoperative short-term outcome and long-term survival were analyzed among the three groups.Results:A total of 96 patients were enrolled. The total laparoscopic group enrolled 29 patients, including 21 males and 8 females, aged (57.8±1.6) years old; the open surgery group enrolled 28 patients, including 18 males and 10 females, aged (57.3±2.0) years old; 39 cases were included in the hybrid surgery group, including 27 males and 12 females, aged (55.3±1.8) years old. The distribution ratio of the two lobes of liver metastases in the open surgery group was higher than that in the total laparoscopic group and hybrid surgery group (all P<0.05), and there was no significant difference in the other clinical baseline characteristics between the three groups (all P>0.05). In laparoscopy group, open surgery group and hybrid surgery group, the mean operative time was (241.5±12.9) min, (209.3±10.7) min and (234.9±12.4) min, respectively. The median intraoperative blood loss was 200.0 ml, 300.0 ml and 200.0 ml, respectively. The median postoperative hospital stay was 8.0 days, 9.0 days and 9.0 days, respectively. There were no statistical differences in these indicators (all P>0.05). The patients in the open surgery group had a longer initial defecation time than those in the other two groups ( P<0.05). The incidence of postoperative complications was 31.0% (9/29), 39.3% (11/28) and 35.9% (14/39), respectively, with no difference among the three groups ( P>0.05). In laparoscopy group, open surgery group and hybrid surgery group, 1-year overall survival were 93.0%, 85.0% and 94.0%; 3-year overall survival were 72.0%, 81.0% and 74.0%, respectively ( P>0.05). One-year disease free survival were 70.0%, 52.0% and 55.0%; 3-year disease free survival were 36.0%, 30.0% and 39.0%, respectively ( P>0.05). Conclusion:Laparoscopic simultaneous resection for left-sided CRC and SLM shows slight advantages in the safety and short-term outcome, and does not affect the long-term survival.

Objective To compare the effects of bone transport versus induced membrane technique for large segmental tibial defects.Methods The clinical data were analyzed retrospectively of 89 patients with large segmental tibial defect who had been treated at Department of Orthopaedics,Wuxi No.9 People's Hospital from June 2005 to February 2017 using bone transport or induced membrane technique.They were 58males and 31 females,aged from 13 to 74 years (average,38.0 years).The bone transport group had 59cases and the induced membrane technique group 30 cases.The 2 groups were compared in terms of preoperative general data and postoperative bone nonunion,bone healing time,complications and functional recovery of the adjacent joint.Results There were no statistically significant differences between the 2groups in terms of age,gender,cause or type of defects,associated injury,course of disease,functionary scores of the adjacent joint or number of operations,showing compatibility between the 2 groups (P ＞ 0.05).All the patients were followed up for 12 to 48 months (average,20 months).The bone transport group had significandy longer clinical healing time (14.7 ± 5.4 months) and significantly higher incidences of major complications (50.8％),minor complications (57.6％) and overall complications (83.1％) than the induced membrane technique group (11.2 ± 2.8 months,16.7％,26.7％ and 30.0％,respectively) (P ＜ O.05),but significantly lower functionary scores of the adjacent joint (86.4 ± 5.0 points) than the induced membrane technique group (88.8 ± 4.9 points) (P ＜ 0.05).Conclusions Both bone transport and induced membrane technique are effective repairs for large segmental tibial defects.However,induced membrane technique may be superior to bone transport in terms of bone healing,complications and functional recovery.

Objective: To evaluate the effects of 4 gingival retraction methods. Methods: 48 premolars were selected from 12 periodontal healthy volunteers and were randomly divided into 4 groups(n = 12 teeth) and treated with different gingival retraction methods: Ultrapak E(group UL) and three kinds of cordless gingival retraction, including Expasy1 (group EX), Astringent Retraction Paste (group AS) and Racegel(group RA). Perfusing plaster models were prapaired after useing putty-wash impression technique. A 3D digital model was reconstructed by a 3-shape model scanner for each patient. Data were collected before and after gingival retraction and analysed by the Geomagic Studio 2013 in the mesial, middle, and distal sides of the selected teeth. Results: Effective width of gingival retraction were all obtained in the 4 groups. The most effective effects were observed in group UL, followed by group EX, group AS and group RA, among the groups, F = 1 114. 4, P＜ 0. 000 1. Conclusion: Cordless gingival retraction technic can achieve effective gingival retraction.

Objective: To investigate the potential effect of proteoglycans (PG) and glycosaminoglycans (GAG) on the stability of resin-dentin bonds against artificial saliva storage. Methods: Seventy-two extracted molars were used to obtain standard dentin bonding surface, and the specimens were etched for 15 s with 37% phosphoric acid and divided into three groups using a table of random number. Then the three groups undergone different incubating procedures as follow: specimens in chondroitinase ABC (C-ABC) group were incubated with C-ABC, specimens in trypsin (TRY) group were incubated with trypsin, and specimens in the control group were incubated with deionized water. All specimens were incubated at 37 ℃ for 48 h in the oscillators. Then specimens in each group were randomly assigned into three subgroups (n=8) as follows: immediate control subgroup, aging subgroups with artificial saliva storage for 6 months and 12 months. Microtensile bond strength (μTBS), fracture mode, bonding interface morphology and nanoleakage were evaluated. Results: Immediately and with artificial saliva storage for 6 months and 12 mouths, the μTBS of TRY group ([49.04±3.57], [37.01±3.21] and [35.27±3.56] MPa) were significantly higher than those in the control group ([40.71±3.32], [28.87±2.34] and [24.20±2.07] MPa) (P<0.05). The immediate μTBS of C-ABC group ([32.94±2.45] MPa) was significantly lower than that of the control group (P<0.05). While with artificial saliva storage for 6 months and 12 mouths, the μTBS of C-ABC group ([26.46±2.45] and [22.50±2.58] MPa) were no differences with those of the control group (P>0.05). The ratio of cohesive fracture increased with the extension of aging time. Some narrow gaps were found in hybrid layer of the control group with artificial saliva storage for 6 months and 12 mouths. Conclusions Removal of PG increased the μTBS and durable bonds to dentin, while removal of GAG decreased the μTBS, however, it can be of help to create more durable bonds to dentin.

Objective:To introduce a new fixation set-up for micro-tensile test.Methods:Dentin-composite were bonded with AdperTM Single Bond 2 (SB2)and sectioned into stick-shaped specimens.Specimens from each tooth(n =6)were equally divided into Ciucchi's jig and the designed set-up(Control and experimental)groups for micro-tensile bond test according to the utilized fixa-tion set-up.The bonding interface failure mode was examined with field-emission scanning electron microscope (FESEM).Three-dimensional models of the two set-ups and the specimen were developed,stress distribution was analyzed by finite element analysis (FEA).Results:The bond strength(MPa)of experimental and control group was 32.76 ±7.43 and 43.58 ±4.72(P <0.05),the ratio of mixed failure was 28 /36 and 20 /36(P <0.05)respectively.FEA showed that the designed set-up for fixing the sticks pro-vided a uniform stress distribution along the long axis of the specimen.FEA and failure mode analysis confirmed such uniform distri-bution was also concentrated at the bonding interface.Conclusion:The new set-up is feasible for micro-tensile test.

Objective To introduce the method of dual immunofluorescence labeling of human dentin matrix without demineralization of the whole dentin fragments, and to analyze the distribution of type-Ⅰ collagen fibrils and chondroitin sulfate in human dentin.Methods Forty 30 μm-thick middle coronal dentin sections were obtained from 8 freshly extracted human third molars and etched with 37％ phosphoric acid(PA) gel for 15 s.After preconditioning with or without tosyl-phenylalanyl chloromethyl ketone(TPCK) treated trypsin digestion, sections were subjected to dual immunofluorescent labeling and scanned by confocal laser scanning microscopy to identify the type-Ⅰ collagen fibrils and chondroitin sulfate.Results Chondroitin sulfate was localized in the lumens of the dentin tubules and peritubular dentin, while the type-Ⅰ collagen fibrils were localized in intertubular dentin and peritubular dentin.After preconditioning with TPCK treated trypsin digestion, the red fluorescence was decreased or disappeared.Conclusions The dual immunofluorescence labeling methodology can be used to study the human dentin matrix without demineralization of the whole dentin fragments.Chondroitin sulfate was localized in the lumens of the dentin tubules and peritubular dentin, while the type-Ⅰ collagen fibrils were localized in intertubular dentin and peritubular dentin.

OBJECTIVETo investigate the differences of clinical characteristics and molecular features among colorectal cancer subsides and provide evidence for colorectal cancer protection, diagnosis and treatment.

METHODSAll of 4 316 colorectal patients from Shanghai cancer center were selected for clinical character analysis, among which, 2 224 subjects for molecular feature analysis. Clinic pathological characteristics like age, gender, tumor types, histological types, differentiation and T-stage, as well as molecular features like hMLH1, hMSH2, CD44, p21, p53, COX2,E-cadherin, Her2 and Ki-67, were involved into this research.

RESULTSIt showed that compared with left-sided colon and rectal cancers, right-sided cancers occurred more in women (46.0% (541/1 176); 39.2% (424/1 083); 41.2% (848/2 057), respectively, χ² = 11.85, P < 0.01), had more mucinous or signet-ring carcinoma (12.0% (128/1 064), 5.8% (56/960), 4.0% (75/1 859), respectively, χ² = 31.27, P < 0.01), poor differentiated carcinoma (32.1% (343/1 069), 19.5% (201/1 033), 19.3% (380/1 967), respectively, χ² = 72.66, P < 0.01) , and advanced T stage (87.9% (992/1 129), 83.2% (869/1 045), 72.2% (1 486/2 057), respectively, χ² = 121.44, P < 0.01). Meanwhile, the rates of hMLH1 were higher in right-sided colon cancers when compared with rectal cancers (13.4% (59/439) vs 8.5% (88/1 035), OR (95%CI): 1.67 (1.18-2.37)), as well as the rates of hMSH2 negative expression (4.9% (22/452) vs 2.4% (26/1 083), OR (95% CI): 2.08(1.17-3.71)). The rates of p53 positive expression were higher in right-sided colon cancers when compared with rectal cancers (76.2% (321/421) vs 68.4%, (776/1 134), OR (95% CI): 0.68 (0.52-0.87)). Compared with right-sided colon cancers, the rates of Her2 positive expression were higher in rectal cancers (19.3% (176/913) vs 13.2% (45/340), OR (95% CI): 1.57 (1.10-2.23)) , as well as the rates of Ki-67 expression which was positive in more than 50% cells (73.6% (840/1 141) vs 65.6% (299/456), OR (95% CI): 0.68 (0.54-0.86)).

CONCLUSIONThere are specific characteristics in right-sided colon cancers. The difference of molecular features between right-sided colon and rectal cancers are more significant.

Adenocarcinoma, Mucinous ; Cadherins ; Carcinoma ; Carcinoma, Signet Ring Cell ; China ; Colonic Neoplasms ; Colorectal Neoplasms ; Female ; Humans ; Neoplasm Grading ; Neoplasm Staging ; Rectal Neoplasms ; Sex Factors

Objective To investigate the differences of clinical characteristics and molecular features among colorectal cancer subsides and provide evidence for colorectal cancer protection,diagnosis and treatment.Methods All of 4 316 colorectal patients from Shanghai cancer center were selected for clinical character analysis, among which, 2 224 subjects for molecular feature analysis.Clinic pathological characteristics like age, gender, tumor types, histological types, differentiation and T-stage, as well as molecular features like hMLH1,hMSH2,CD44,p21,p53,COX2,E-cadherin,Her2 and Ki-67,were involved into this research.Results It showed that compared with left-sided colon and rectal cancers, right-sided cancers occurred more in women ( 46.0% ( 541/1 176 ); 39.2% ( 424/1 083 ); 41.2% ( 848/2 057 ) , respectively,χ2 =11.85,P<0.01), had more mucinous or signet-ring carcinoma(12.0%(128/1 064), 5.8%(56/960),4.0%(75/1 859),respectively,χ2 =31.27,P <0.01), poor differentiated carcinoma (32.1%(343/1 069),19.5%(201/1 033), 19.3%(380/1 967),respectively,χ2 =72.66,P<0.01), and advanced T stage(87.9%(992/1 129), 83.2%(869/1 045), 72.2%(1 486/2 057),respectively,χ2 =121.44,P <0.01).Meanwhile,the rates of hMLH1 were higher in right-sided colon cancers when compared with rectal cancers ( 13.4% ( 59/439 ) vs 8.5% ( 88/1 035 ) , OR ( 95%CI ): 1.67 ( 1.18-2.37)),as well as the rates of hMSH2 negative expression(4.9% (22/452) vs 2.4% (26/1 083),OR (95%CI):2.08(1.17-3.71)).The rates of p53 positive expression were higher in right-sided colon cancers when compared with rectal cancers ( 76.2% ( 321/421 ) vs 68.4%, ( 776/1 134 ) , OR ( 95%CI ): 0.68 (0.52-0.87)).Compared with right-sided colon cancers,the rates of Her2 positive expression were higher in rectal cancers(19.3%(176/913) vs 13.2% (45/340),OR(95%CI):1.57 (1.10-2.23)),as well as the rates of Ki-67 expression which was positive in more than 50%cells (73.6%(840/1 141) vs 65.6%(299/456),OR(95%CI):0.68 (0.54-0.86)).Conclusion There are specific characteristics in right-sided colon cancers.The difference of molecular features between right-sided colon and rectal cancers are more significant.