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.

Pyroptosis is a kind of inflammatory cell death mediated by the Gasdermin family,which has made great progress in tumor therapy in recent years.Basing on that cell pyroptosis can activate the anti-tumor immune response,and tumor immunotherapy is a new field of tumor therapy,the regulation of cell pyroptosis exhibits great potential for tumor therapy.Meanwhile,nanotechnology is the key means of tumor precision treatment with the advantages of precise targeting and continuous release.Based on these current situations,this paper summarizes the drugs that activate pyroptosis and the nanocarriers that use nanotechnology to promote pyroptosis to participate in tumor therapy,and summarizes the mechanism and application of their action on pyroptosis.This paper is aimed to provide certain references for anti-tumor therapy based on pyroptosis.

Objective:To investigate the risk factors of acute gastrointestinal failure in patients with traumatic brain injury(TBI).Methods:A retrospective analysis of 297 patients admitted to the Rugao Hospital Affiliated to Nantong University for traumatic brain injury from March 2018 to March 2020 was performed. Multivariate Logistic regression analysis was used to test the risk factors of acute gastrointestinal failure in patients with traumatic brain injury.Results:Of the 297 patients with TBI, 92 (30.98%) had acute gastrointestinal failure, of which 56 (18.86%) were acute gastrointestinal injury(AGI) Ⅲ and 36 (12.12%) were AGI Ⅳ. Multivariate Logistic regression analysis showed that female, low Glasgow Coma Scale (GCS) scores, frontal lobe injury, abnormal serum sodium, lung infection and intracranial infection were independent risk factors for acute gastrointestinal failure in TBI patients ( P<0.05). The length of ICU stay in the acute gastrointestinal failure group was significantly higher than that of the non-acute gastrointestinal failure group: (18.5 ± 2.6) d vs. (6.3 ± 1.2) d, and the incidence of good prognosis was significantly lower than that of the non-acute gastrointestinal failure group: 60.87%(56/92) vs. 80.49(165/205), the differences were statistically significant ( P<0.05). Conclusions:That female, low GCS scores, frontal lobe injury, abnormal serum sodium, lung infection and intracranial infection are independent risk factors for acute gastrointestinal failure in TBI patients

Objective: To explore the efficacy and safety of recombinant human interleukin-11 (rhIL-11) in treatment of chemotherapy-induced thrombocytopenia of acute leukemia. Methods: Acute leukemia patients with chemotherapy-induced thrombocytopenia [Platelets (Plt) < 50×10⁹/L] in 6 centers nationwide from February 2016 to July 2016 were treated with rhIL-11 (2 mg/time, twice per day) by subcutaneous injection. Treatment lasted 7 days or at least until Plt≥ 50×10⁹/L. The Plt recovery was observed during treatment. Results: A total of 112 patients were enrolled, and 2 patients decided to drop out of study. The efficacy population consisted of 110 patients, and the total response rate reached 74.5% (82/110). The average variation of Plt during treatment was (70±54)×10⁹/L, and recovery average time of Plt for the patients with favorable efficacy was (8.7±3.0) days. In treatment with severe thrombocytopenia, rhIL-11 alone could shorten the recovery time compared with rhIL-11 combined with Plt transfusion [(8.0±2.6) d vs. (9.6±3.5) d, t=2.17, P=0.03]. Conclusion rhIL-11 twice a day of subcutaneous injection can effectively promote Plt recovery and reduce Plt transfusion with less adverse reactions, which is worthy of further application.

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.

Objective To investigate the effect of four different cements on the bonding effectiveness of root canal dentine and fiber post before and after different treatments.Methods A total of 216 freshly extracted sound single-root-canal mandibular premolars were randomly divided into four groups.After root canal treatment and post space preparation being conducted on the premolars,Fuji Ⅰ,Fuji Cem,RelyX Unicem,RelyX ARC were used respectively to bond fiber posts and were marked with group A,B,C,and D.Microleakage,micromorphology of the bonded interfaces,and pull-out bond strength were evaluated in the immediate group,thermocycling group and thermomechanical loading group.Results In the immediate group,samples in group D showed the highest bond strength [(278 ± 26) N],followed by group C[(219 ±12) N],B[(104 ±23) N] and A[(73 ±8) N].Significant differences were found among all groups (P ＜ 0.05).A significant increase in bond strength was found in group A and B,whereas a decrease tendency was detected in group C and D after different treatments.Scanning electron microscope indicated that some little gaps were observed in group D after treatment,while a more intense bonding interface was found in group A and B.Microleakage scores in group A and B were lower than those in group C and D after aging treatments.Conclusions Resin cement can achieve a better immediate bond strength,while resinmodified resin cement may acquire a better long-term retention.

Objective To investigate the potential effect of proteoglycan(PG) and glycosaminoglycan(GAG) on the bonding of etch and rinse adhesive to dentin,in order to improve the bonding effect of dentin.Methods Forty-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 numbers.Then the three groups undergone different incubating procedures as follow:specimens in chondroitinase ABC(C-ABC) group were incubated with C-ABC at 37 ℃ for 48 h in vibrator.Specimens in trypsin(TRY) group were incubated with trypsin,and specimens in the control group were incubated with deionized water for 48 h in the oscillators.Then specimens in each group were randomly assigned into two subgroups,A(AdperTM Single Bond 2) and B(Prime & Bond NT) (n=7).The microtensile bond strength(μTBS),fracture mode and bonding interface morphology of the specimens were evaluated via microtensile testing,stereo microscope and field emission scanning electron microscopy(FE-SEM) respectively after specimens being incubated in 37 ℃ water for 24 h.Results The immediate μTBS of C-ABC group bonding with adhesive A and B [(32.9±2.5) and (26.8±2.2) MPa] were significantly lower than that of the control group [(40.7±3.3) and (34.6±3.7) MPa](P＜0.05).While the immediate μTBS of TRY group [(49.0± 3.6) and (44.5± 3.0) MPa] were significantly higher than that of the control group(P＜0.05).Conclusions Dentin PG participates in the dentin bonding process.Removal of PG increased the immediate μTBS of dentin and total etching adhesives,while removal of GAG decreased the immediate μTBS.

Objective To evaluate the colonoscopy surveillance in colorectal cancer patients after surgical removal of the tumor.Methods From June 1986 to June 2007,2762 asymptomatic patients who had underwent operation for colorectal cancer were put into colonoscopy surveillance.They had the first examination 3-6 months after the operation,and were re-examined once a year thereafter for 3 years.The follow-up findings were compared with those from the 218 symptomatic patients who had colorectal cancer surgery from September 1981 to May 1986.Results In 2762 asymptomatic patients,48 cases of multiple primary cancer were detected,in which 39 cases(1.4%) were found at one examination and 9 cases(0.3%)at different examination.The TNM staging of these lesions included stage Ⅰ in 6,stage Ⅱ in 31 and stage Ⅲ in 11.During the surveillance,583 cases(21.1%) of adenoma were detected and endoscopically resected,in which 17(3.2%) were invasive early cancer and 58(9.9%) were high grade dysplasia.In 218 patients with symptoms,29 cases(13.3%) of adenoma and 27 cases( 12.4%) of cancer were detected,including 4 cases of stage Ⅰ cancer,6 of stage Ⅱ and 16 of stage Ⅲ.Conclusion Colonoscopy surveillance in colorectal cancer patients after surgery is important in finding precancerous lesion and early stage cancer,and is recommended in all patients.