AIM: To study the roles of Toll-like receptor 4 ( TLR4 ) and TLR4 activator lipopolysaccharide ( LPS) in colonic inflammation recovery .METHODS:Normal intestinal epithelial cells were cultured with LPS in vitro. The subgroups of the intestinal epithelial cells with differential expression of TLR 4 ( low, normal and high ) were construc-ted by the technique of lentivirus transfection .The cells with normal and high expression of TLR 4 were induced by LPS for 0 h, 2 h and 4 h.Inflammatory cytokines TNF-α, IL-6 and IL-8 in the culture supernatant were detected by ELISA .The mRNA levels of TNF-α, IL-6, IL-8, IL-10 and IL-1βwere detected by qPCR .The cell mobility was also monitored by wound healing assay .RESULTS:The protein expression of TLR 4 was significantly higher after LPS treatment than that in control groups of both cells with TLR4 normal and high expression (P<0.05).The inflammatory cytokines TNF-α, IL-6, IL-8 and IL-1βat mRNA and protein levels were also significantly increased after LPS treatment compared with control group (P<0.05).The protein levels of TNF-α, IL-6 and IL-8 between the 2 groups were also different with statistical sig-nificance ( P<0.05 ) .Higher mobility was observed in the cells with TLR 4 high expression compared to control cells . CONCLUSION:LPS induction might play a role in the activation of TLR 4-mediated inflammatory pathways by up-regula-ting the expression of inflammatory cytokines at both transcriptional and translational levels .

Objective To investigate the relationship between acute radiation proctitis and radiation dose,volume as well as radiation time,in the process of intensity-modulated radiation therapy (IMRT) for the cervical cancer patients.Methods A total of 51 patients with locally advanced cervical cancer were enrolled from January 2011 to December 2013.Those patients were then classified into grade 1 to 4 groups,according to the RTOG/EORTCtoxicity grading standard.The exposure dose volume and the average dose of rectum under the standard plan were evaluated with dose-volume histogram (DVH).The ANOVA test was used for analyzing Dmax,D mean,D1 cm3,D2cm3,D40 and V40 values of rectum and the average exposure dose of rectum.Results The average time of acute radiation proctitis with clinical symptoms was (23.06 ± 12.01) d after radiotherapy.Dmaxvalues of rectum in grade 2 group was lower than those in grade 3 and 4 groups (F =5.268,P ＜ 0.05).Moreover,D1 cm3 and D2 cm3 values of rectum in grade 1 and 2 groups were also lower than those in grade 3 and 4 groups (F =4.893,4.406,P ＜ 0.05).There was no statistically significant difference between D40 and V40 values.Conclusions The acute radiation proctitis could be frequently found around 20 days during the IMRT for cervical cancer patients.Mild and moderate acute radiation proctitis are more common,while severe acute radiation proctitis is rare.Minimizing Dmax,D1 cm3 and D2 cm3 values of rectum might reduce the incidence of severe acute radiation proctitis in cervical cancer patients receiving IMRT.

Objective: To evaluate the efficacy, toxicity and cosmetic outcomes of hypofractionated simultaneous integrated boost intensity-modulated radiotherapy (IMRT-SIB) after breast conservative surgery (BCS) for early breast cancer patients. Methods: A total of 76 patients with stage TisT_1-2N₀M₀ breast cancer treated with BCS were enrolled in the analysis. The patients who underwent breast radiotherapy without regional lymph node irridiation and hypo fractionated IMRT/VMAT were used. All patients received whole breast IMRT/VMAT with tumor bed SIB. The doses delivered to the whole breast was 42.4 Gy in 16 fractions, and the dose delivered to tumor bed for SIB was 49.6 Gy in 16 fractions. Cosmetic evaluation was based on the Harvard system. Acute and late toxicities were scored according to CACAT version 3.0. Survival and recurrence rates were calculated by Kaplan-Meier method. The univariate and multivariate analysis were conducted with logistic regression. Results: The median follow-up was 29 months (range 16-40 months). The follow-up rate was 100%. The 1-, 2-and 3-year overall survival rates were 100%. No recurrence or metastasis was observed in this study. The incidence of grade 1 acute skin toxicity was 68.4%, grade 2 was 7.9%. The late skin toxicity of grade 1 was 13.1%, grade 2 was 2.6%.In all, 82.4% of patients had excellent and good cosmetic outcome. The Mean dose of the tumor bed was predictive factor for grade 2 dermatitis. Conclusion: The efficacy, cosmetic effect, the acute and late treatment-related toxicity of hypofractionated IMRT/VMAT-SIB in patients with early breast cancer following BCS might be acceptable. A longer follow-up is needed to define the efficacy on outcomes. Trial registration Chinese clinical trial registry, ChiCTR1800016287

Ischemia reperfusion (I/R) injury is caused by ischemic shock, cardiac arrest, resection and transplantation, and its mechanism is closely related to calcium overload. Mitochondrial calcium uniporter (MCU) is a highly selective calcium channel located in the mitochondrial inner membrane (IMM), mediating calcium ions into the mitochondrial matrix. The MCU complex with the MCU as the core plays an important role in maintaining calcium ion homeostasis and alleviating I/R injury in the heart, kidney, brain, liver and other organs. The mitochondria associated endoplasmic reticulum membranes (MAMs) is a key protein in this process.

Aim To investigate the role of cholesterol in the regulation of lipid raft and the function of platelets.Methods Using in vitro incubation of methyl β-cyclodextrin(MβCD)and in vivo elevation of peripheral blood total cho-lesterol levels to remove and load platelet cholesterol,respectively.Cholera toxin B staining combined with flow cytometry was used to detect platelet lipid raft content,fluorescence antibody staining combined with flow cytometry was used to detect the expression levels of P-selectin and activated integrin α Ⅱ bβ3,annexin Ⅴ labeling combined with flow cytometry was used to detect the level of phospholipid efflux,in vitro experimental system and rat tail bleeding experiment were used to detect platelet aggregation ability.Results The content of lipid raft on B lymphocytes decreased with the removal of cholesterol,while in vitro incubation of MβCD to remove platelet cholesterol significantly increased its lipid raft level(P＜0.05).Consistent with this,in vivo cholesterol loading increased the lipid raft content of B lymphocytes but decreased the lipid raft content of platelets(P＜0.05).The increase in lipid raft after removing cholesterol was not conducive to platelet activation and aggregation function.In vivo cholesterol loading downregulated platelet lipid raft content(P＜0.05),enhanced its ability to respond to low concentration stimulant for activation aggregation and coagulation,and this enhancing effect disappeared after cholesterol removal.Conclusion Platelet cholesterol is a key regulator of platelet lipid raft content and platelet function,which can negatively regulate lipid raft,promote platelet activation,and enhance their coagulation function.

Objective:To evaluate the role of ubiquitin-specific peptidase 22 (USP22) in myocardial ischemia-reperfusion (I/R) injury in diabetic mice.Methods:Seventy-eight SPF male C57BL/6 mice, aged 6-8 weeks, were divided into 6 groups using a random number table method: sham operation group (Sham group, n=12), type 1 diabetes mellitus + sham operation group (T1D+ Sham group, n=12), myocardial I/R injury group (I/R group, n=12), type 1 diabetes mellitus + myocardial I/R injury group (DI/R group, n=12), type 1 diabetes mellitus + myocardial I/R injury + empty vector group (DI/R+ V group, n=15), and type 1 diabetes mellitus + myocardial I/R injury + USP22 overexpression group (DI/R+ U group, n=15). Type 1 diabetes mellitus was induced by intraperitoneal injection of streptozotocin-citrate buffer. Myocardial I/R was induced by ligation of the left coronary artery. At 1 day before developing the myocardial I/R injury model, DI/R+ U group and DI/R+ V group received an intramyocardial injection of USP22 overexpression plasmid or empty vector plasmid, respectively. At 24 h of reperfusion, cardiac function was assessed using the echocardiography to measure the left ventricular ejection fraction and left ventricular fractional shortening. The mice were then sacrificed, and their hearts were harvested for measurement of the myocardial infarct size, for microscopic examination of pathological changes (using HE staining) and for determination of the apoptosis rate (TUNEL staining), reactive oxygen species(ROS) activity (DHE staining), and USP22 expression (by Western blot, immunofluorescence, and immunohistochemistry). Proteomic analysis was performed to identify downstream proteins regulated by USP22, and protein-protein interactions were investigated using co-immunoprecipitation. Results:Compared with Sham group, the cardiac function indices were significantly decreased, the apoptosis rate of myocardial cells and ROS activity were increased, and USP22 expression in myocardial tissues was down-regulated in I/R group ( P<0.05). Compared with I/R group, the percentage of myocardial infarct size was significantly increased, the cardiac function indices were decreased, the apoptosis rate of myocardial cells and ROS activity were increased, and USP22 expression in myocardial tissues was up-regulated ( P<0.05), and the pathological damage to myocardial tissues was aggravated in DI/R group. Compared with DI/R+ V group, the percentage of myocardial infarct size was significantly decreased, the cardiac function indices were increased, the apoptosis rate of myocardial cells and ROS activity were decreased, and USP22 expression in myocardial tissues was up-regulated ( P<0.05), and the pathological damage to myocardial tissues was alleviated in DI/R+ U group. The results of proteomics combined with co-immunoprecipitation experiments showed an interaction between calponin 1 and USP22. Conclusions:During myocardial I/R injury in diabetic mice, USP22 may act as an endogenous protective mechanism, and calponin 1 might be a downstream mechanism through which USP22 exerts its protective effects.

OBJECTIVE：To analyze the chemotypes of volatile components from Perillae Folium of different germplasms ，and to investigate the relationship of germplasm and leaf color with chemotype. METHODS ：The fingerprints of volatile components from 30 batches of Perillae Folium were prepared by GC-MS with P 4 peak as reference. Similarity Evaluation System for TCM Chromatographic Fingerprint （2004A edition ）was applied to evaluate the similarity and confirm common peaks. The volatile components of Perillae Folium were determined by the same GC-MS method. Qualitative Navigator （B.08.00）software was used to analyze and compare with NIST 17.0 standard mass spectrum database. The compounds corresponding to the peak were analyzed ； clustering analysis was carried out with Origin 2018 software. RESULTS ：There were 13 common peaks in the fingerprints of volatile components from 30 batches of Perillae Folium . The similarities were 0.13-1.00. Totally 54 components were identified from 30 batches of Perillae Folium of different germplasm. Cluster analysis showed that 30 batches of Perillae Folium samples could be clustered into three categories ；among them ，SCY-1，YNT-9，YNX-17，YN-28 were clustered into one category ，with phenylpropanoid-elemicin（PP-e as ）the main volatile component ，being PP-e type ；GS-4，GS-7，GS-11，GS-19，HBA-14， HBA-20，GZZ-8，LN-39，GSL-27，GSQ-32，GSQ-33，GST-31，YNW-12，LN-38 were clustered into one category ，and the content of perilla ketone （PK）in them was the highest except for LN- 38， being PK type [the content of phenylpropanoid-apiol（PP-a）in LN- 38 was higher than that of perilla ketone ，being PP-a type] ；HBS-2，HBS-3，HBS-6， C201859）HBS-15，HBS-16，HBS-24，HBS-25，GX-26，SXS-30，SCC- 36，RB-37，SC-29 were clustered into one category ，and thecontent of perillaldehyde （PA）was the highest ，being PA type.The color characteristics of Perillae Folium of different germplasm showed that Perilla frutescens （L.） Britt. var.frutescens with green leaves on both sides was PK type ，while P. frutescens （L.）Britt. var. arguta with purple leaves on one or both sides was PA type ，and P. frutescens （L.） Britt var. auriculato-dentata C. Y. Wu et Hsuan ex H. W. Li was PP-e type. CONCLUSIONS：The chemotype of volatile components in Perillae Folium have a certain corresponding relationship with their leaf colors. Most of P. frutescens （L.）Britt. var. arguta with purple leaves on one side or both sides are PA type. P. frutescens （L.） Britt. var. acuta （Thunb.）Kudo，P. frutescens （L.）Britt var. auriculato-dentata C. Y. Wu et Hsuan ex H. W. Li and P. frutescens （L.）Britt. var. frutescens with green leaves on both sides do not belong to PA type ，among which P. frutescens （L.）Britt var. frutescens is PK type ，while P. frutescens （L.）Britt var. auriculato-dentata C. Y. Wu et Hsuan ex H. W. Li is mostly PP-e type.