ObjectiveTo investigate the mechanism of Atractylodis Macrocephalae Rhizoma（AMR） in the treatment of slow-transmission constipation（STC） by observing the effects of AMR on short-chain fatty acids and intestinal barries in STC mice. MethodForty-eight male KM mice were randomly divided into blank group， model group， AMR low-， medium-， high-dose groups（2.5， 5， 10 g·kg^-1） and mosapride group（2.5 mg·kg^-1）. Except for the blank group， all groups were gavaged with loperamide suspension（5 mg·kg^-1） twice daily for 14 d to construct the STC mouse model. At the same time， each drug administration group was given the corresponding drug by gavage for consecutive 14 d， the blank and model groups were gavaged with equal volume of distilled water. The effects of the treatment of AMR on body mass， defecation frequency， fecal water content and intestinal propulsion rate of mice were observed， the pathological changes of mouse colon were observed by hematoxylin-eosin（HE） staining and periodic acid-Schiff（PAS） staining， the levels of gastrin（GAS） and motilin（MTL） in serum were detected by enzyme-linked immunosorbent assay（ELISA）， gas chromatography-mass spectrometry（GC-MS） was used to detect the contents of short-chain fatty acids（SCFAs） in mouse feces， real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） and Western blot were used to determine the mRNA and protein expression levels of zonula occludens-1（ZO-1）， Occludin， and Claudin-1 in the colon of mice. ResultCompared with the blank group， the body mass， defecation frequency， fecal water content and intestinal propulsion rate of mice in the model group were significantly decreased（P<0.05， P<0.01）， the arrangement of colonic tissues was disordered， and the number of goblet cells was reduced， the levels of GAS and MTL in serum were significantly decreased（P<0.01）， and the levels of SCFAs in the feces were on a decreasing trend， with the contents of acetic acid， propionic acid， butyric acid， isobutyric acid and valeric acid were significantly decreased（P<0.05， P<0.01）， the mRNA and protein expression levels of ZO-1， Occludin and Claudin-1 in the colonic tissues were significantly decreased（P<0.01）. The above results suggested that STC mouse model was successfully constructed. Compared with the model group， the body mass， defecation frequency， fecal water content and intestinal propulsion rate of mice in AMP administration groups all increased significantly（P<0.05， P<0.01）， the mucosal layer of the colonic tissues was structurally intact without obvious damage， and the number of goblet cells increased， serum levels of GAS and MTL were significantly increased（P<0.01）， the contents of SCFAs in the feces were all on a rising trend， with the contents of acetic， propionic， butyric and isobutyric acids rising significantly（P<0.05， P<0.01）， the mRNA and protein expression levels of ZO-1， Occludin and Claudin-1 in the colonic tissues were significantly increased（P<0.05， P<0.01）. ConclusionAMR is able to improve the constipation symptoms in STC mice， and its mechanism may be related to increasing the contents of SCFAs in the intestine as well as promoting the mRNA and protein expression levels of ZO-1， Occludin and Claudin-1 in the colon.

BACKGROUND:Tissue engineering has brought new hope to the clinical challenge of liver failure,and the preparation of plant-derived decellularized fiber scaffolds holds significant importance in liver tissue engineering. OBJECTIVE:To prepare apple tissue decellularized scaffold material by using fresh apple slices and a solution of sodium dodecyl sulfate,and assess its biocompatibility. METHODS:Fresh apples were subjected to decellularization using phosphate buffer saline and sodium dodecyl sulfate solution,separately.Afterwards,the decellularized apple tissues and apple decellularized scaffold materials were decontaminated with phosphate buffer saline.Subsequently,scanning electron microscopy was used to assess the effectiveness of decellularization of the apple materials.Adipose-derived mesenchymal stem cells were extracted from the inguinal fat BALB/C of mice,and their expression of stem cell-related markers(CD45,CD34,CD73,CD90,and CD105)was identified through flow cytometry.The cells were then divided into a scaffold-free control group and a scaffold group.Equal amounts of adipose-derived mesenchymal stem cells were seeded onto both groups.The biocompatibility of the decellularized scaffold with adipose-derived mesenchymal stem cells was evaluated using CCK-8 assay,hematoxylin-eosin staining,and phalloidine staining.Cell adhesion and growth on the scaffold were observed under light microscopy and scanning electron microscopy.Furthermore,the scaffold was subdivided into the non-induced group and the hepatogenic-induced group.Adipose-derived mesenchymal stem cells were cultured on the decellularized apple scaffold,and they were cultured for 14 days in regular culture medium or hepatogenic induction medium for comparison.Immunofluorescent staining using liver cell markers,including albumin,cytokeratin 18,and CYP1A1,was performed.Enzyme-linked immunosorbent assay was used to detect the secretion of alpha fetoprotein and albumin.Additionally,scanning electron microscopy was employed to observe the morphology of the induced cells on the scaffold,verifying the expression of liver cell-related genes on the decellularized scaffold material.Finally,the cobalt-60 irradiated and sterilized decellularized apple scaffolds were transplanted onto the surface of mouse liver and the degradation of the scaffold was observed by gross observation and hematoxylin-eosin staining after 28 days. RESULTS AND CONCLUSION:(1)The scanning electron microscopy results revealed that the decellularized apple scaffold material retained a porous structure of approximately 100 μm in size,with no residual cells observed.(2)Through flow cytometry analysis,the cultured cells were identified as adipose-derived mesenchymal stem cells.(3)CCK-8 assay results demonstrated that the prepared decellularized apple tissue scaffold material exhibited no cytotoxicity.Hematoxylin-eosin staining and phalloidine staining showed that adipose-derived mesenchymal stem cells were capable of adhering and proliferating on the decellularized apple tissue scaffold.(4)The results obtained from immunofluorescence staining and enzyme-linked immunosorbent assay revealed that adipose-derived mesenchymal stem cells cultured on the decellularized apple scaffolds exhibited elevated expression of liver-specific proteins,including albumin,alpha-fetoprotein,cytokeratin 18,and CYP1A1.These results suggested that they were induced differentiation into hepatocyte-like cells possessing functional characteristics of liver cells.(5)The decellularized apple scaffold implanted at 7 days has integrated with the liver,with partial degradation of the scaffold observed.By 28 days,the decellularized apple scaffold has completely degraded and has been replaced by newly-formed tissue.(6)The results indicate that the decellularized scaffold material derived from apple tissue demonstrates favorable biocompatibility,promoting the proliferation,adhesion,and hepatic differentiation of adipose-derived mesenchymal stem cells.

Background Working night shifts is common in the secondary industry workers, and usually with long weekly working hours. They are prone to occupational stress and sleep disorders, which may seriously affect their physical and mental health and work efficiency. Objective To investigate the current situation and influencing factors of occupational stress and sleep disorders among three key occupational groups in Guizhou Province, and provide a basis for formulating intervention measures. Methods A stratified random cluster sampling method was used to select 11552 workers as the research subjects from three occupational groups in Guizhou Province. A survey on occupational stress and sleep disorders was conducted using the National Key Population Occupational Health Literacy Monitoring Survey Personal Questionnaire. Results A total of 9956 valid questionnaires were recovered (86.18%). The positive rates of occupational stress and sleep disorders among the three key occupational groups in Guizhou Province were 22.6% and 40.6%, respectively. No differences were found among those by individual characteristics (P>0.05). There was a statistically significant difference in reporting sleep disorders among the occupational groups by industry (P<0.05), with the highest rate observed in the production and supply industries of electricity, heat, gas, and water (47.9%). The results of logistic regression analysis showed that company size, weekly working hours, sleep disorders, length of service, night shift, monthly income, and gender were all independent influencing factors of occupational stress in the target population (P <0.05); occupational stress, weekly working hours, marital status, industry, educational level, night shift, household type, monthly income, and age were all independent influencing factors for sleep disorders in the target population (P <0.05). Conclusion The positive rates of occupational stress and sleep disorders are high among the three key occupational groups in Guizhou Province, with sleep disorders particularly prominent in the production and supply industries of electricity, heat, gas, and water. Special attention should be paid to sleep disorders in individuals of the industry of the production and supply industries of electricity, heat, gas, and water, under 40 years old, widowed or divorced, with low education, low income, working night shifts, long weekly working hours, and experiencing occupational stress.

Objective To observe the neuropsychiatric behavioral performance of kainic acid(KA)-induced epilepsy rats;investigate gender differences in acute seizure and behavioral performance tasks relating to sense,motor,learning,and memory in the remission phase;and explore the potential neurobiological mechanisms of action.Methods Healthy SD rats aged 4 weeks were randomly divided into control and model groups,with 22 rats in each group(11 males and 11 females).An epileptic rat model was induced by intraperitoneal injection of KA.Seizure latency and frequency within 2 hours of KA injection were observed,seizure grade was assessed using the Racine grade standard,and a cortical electroencephalogram(EEG)was recorded.Behavioral performance was observed in a series of tasks including open field testing,balance beam walking,elevated plus maze,Y-maze,and novel object recognition.The level of GABA in the hippocampus was detected by ELISA,injury to hippocampal neurons was observed by Nissl staining,and the protein expression of synapsin-1 and synaptotagmin 1 in the hippocampus were detected by Western Blot.Results Both male and female rats presented typical epileptic behaviors after KA injection.However,compared with the effects in males,the latency of the first seizure(P=0.014)and Ⅳ～Ⅴ grading in female model rats were more pronounced(P＜0.01),and the frequency of epileptic seizures within 2 hours was significantly reduced(P=0.019).In the open field testing,KA-induced epileptic rats presented more motor but fewer hedonic behaviors,as indicated by the decrease in total movement distance in the central area,compared with the control group.Moreover,grooming frequency was significantly reduced in the female model rats compared with not only that in the control but also that in male model rats(P＜0.01).The model rats spent more time completing and had a higher score in the balance beam walking task,indicating their poorer stability and balance.In the elevated plus maze,the exploration times of male model rats in the closed arm was increased.The preference index of rats for the novel arm or object decreased in the Y-maze and novel object recognition,suggesting impairments to their learning and memory abilities.Moreover,neuronal injuries were found in the hippocampus of the model rats that were accompanied with a decline in GABA concentration and protein expression of synapsin-1 and synaptotagmin 1,with no gender differences.Conclusions Intraperitoneal injection of KA successfully induced an epilepsy rat model.However,there was a gender difference in the characters of acute seizures and performance of sensory,motor,and learning memory during epileptic remission.There was no gender differences in the hippocampal GABA concentration or expression of synaptic plasticity-related proteins,and thus no evidence was found for the mechanisms underlying the gender differences.

Objective To investigate how the neutrophil extracellular traps (NETs) affect the proliferation and migration of mouse MC38 colorectal cancer cells and its mechanism. Methods Spleen neutrophils were extracted in mouse, followed by collection of NETs after ionomycin stimulation in vitro. The proliferation of MC38 cell was detected by CCK-8 assay, and migration ability were detected by Transwell^TM and cell scratch assay, after co-incubation with MC38 cells. The mRNA expression of cellular matrix metalloproteinase 2 (MMP2) and MMP9 were detected by real-time fluorescence quantitative PCR, and the expression of MMP2, MMP9 and focal adhesion kinase (FAK), phosphorylated FAK protein were detected by Western blot. After silencing MMP9 using small interfering RNA (siRNA), the effect of NETs on the proliferation and migration ability of MC38 cells and the altered expression of related molecules were examined by previous approach. Results NETs promoted the proliferation and migration of MC38 cells and up-regulated the MMP9 expression and FAK phosphorylation. Silencing MMP9 inhibited the promotion of MC38 proliferation and migration by NETs and suppressed FAK phosphorylation. Conclusion NETs up-regulates MMP9 expression in MC38 cells, activates FAK signaling pathway and promotes tumor cell proliferation and migration.
Animals ; Mice ; Focal Adhesion Protein-Tyrosine Kinases/metabolism* ; Matrix Metalloproteinase 2/metabolism* ; Matrix Metalloproteinase 9/metabolism* ; Extracellular Traps/metabolism* ; Cell Movement ; Cell Proliferation ; RNA, Small Interfering/genetics* ; Colorectal Neoplasms/genetics* ; Cell Line, Tumor

Articular cartilage (AC) injuries often lead to cartilage degeneration and may ultimately result in osteoarthritis (OA) due to the limited self-repair ability. To date, numerous intra-articular delivery systems carrying various therapeutic agents have been developed to improve therapeutic localization and retention, optimize controlled drug release profiles and target different pathological processes. Due to the complex and multifactorial characteristics of cartilage injury pathology and heterogeneity of the cartilage structure deposited within a dense matrix, delivery systems loaded with a single therapeutic agent are hindered from reaching multiple targets in a spatiotemporal matched manner and thus fail to mimic the natural processes of biosynthesis, compromising the goal of full cartilage regeneration. Emerging evidence highlights the importance of sequential delivery strategies targeting multiple pathological processes. In this review, we first summarize the current status and progress achieved in single-drug delivery strategies for the treatment of AC diseases. Subsequently, we focus mainly on advances in multiple drug delivery applications, including sequential release formulations targeting various pathological processes, synergistic targeting of the same pathological process, the spatial distribution in multiple tissues, and heterogeneous regeneration. We hope that this review will inspire the rational design of intra-articular drug delivery systems (DDSs) in the future.

Objective:To analyze the expression and clinical significance of reticulocalbin 3 (RCN3) in colon cancer by bioinformatics database and biological experiments.Methods:Colon cancer HT29 and SW620 cells and colon normal mucosal cells FHC were cultured. The expression of RCN3 in cells was verified by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) and Western blot. The expression data of RCN3 in normal colon tissue and colon cancer tissue were obtained by Ualcan database. The co-expressed gene information of RCN3 from LinkedOmics database was obtained, and the biological processes and related functions of these RCN3 co-expressed genes through were analyzed by gene ontology analysis (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. The protein-protein interaction network of RCN3 related coding genes was constructed by using STRING database. Finally, the relationship between the expression of RCN3 and the clinical prognosis of patients with colon cancer was compared and analyzed according to GEPIA, Ualcan and Linked Omics biological database.Results:Western blot and qRT-PCR results showed that the mRNA and protein expression of RCN3 in HT29 and SW620 colon cancer cells was significantly higher than those in FHCcells ( all P<0.05). The analysis of biological database showed that the expression level of RCN3 in colon cancer tissue was higher than that in normal colon tissue ( P<0.05). GO enrichment analysis showed that RCN3 co-expression genes were mainly involved in the composition of extracellular matrix and extracellular domain structure, the binding process of extracellular matrix and multiple receptors, and the biological processes related to tumor development such as cell adhesion, immune response, and angiogenesis through extracellular domain structure. KEGG pathway analysis showed that RCN3 co-expression genes were mainly involved in ECM receptor interaction, cytokine receptor interaction, chemokine signaling pathway, phosphatidylinositol-3-kinase protein kinase B (PI3K-Akt) signaling pathway, phagosome signal, IgA related intestinal immune network signal, these signaling pathways always related to tumor invasion, migration and inflammatory immune response. The protein-protein interaction network analysis showed that the coding protein genes that directly interacted with RCN3 protein that included PRDX6, NOSIP, PCSK6, IMMP1L, PRRG2, FBXO47, FCGRT, FKBP9, PCDHGA12, and PNMAL1, which were mainly involved in the occurrence and development of colorectal cancer, liver cancer, gastric cancer, breast cancer, lung cancer, and ovarian cancer. Survival curve analysis showed that the overall survival rate of colon cancer patients with high expression of RCN3 was significantly lower than that of patients with low expression of RCN3 ( P<0.05). Conclusions:RCN3 is highly expressed in colon cancer tissues and cells, which is closely related to the occurrence, development and prognosis of colon cancer. It can be used as one of the markers for early screening and prognosis prediction of colon cancer.

Objective:To master the epidemic status and characteristics of skeletal fluorosis in the coal-burning-borne fluorosis affected areas of Chongqing, and to provide a scientific basis for formulating accurate prevention and control strategy for elimination of coal-burning-borne fluorosis.Methods:Stratified sampling method was used to select the villages with mild, moderate and severe coal-burning-borne fluorosis in Wushan and Pengshui, respectively in January-November 2018. The number of villages surveyed in each area was determined by the proportion of 5% to 10% of the actual number of the diseased villages. To investigate the resident population, all the adults over 25 years old in the village were examined for skeletal fluorosis through clinical and X-ray examination, and were diagnosed according to the "Diagnostic Criteria of Endemic Skeletal Fluorosis" (WS 192-2008). The prevalence of skeletal fluorosis in different disease areas, different sexes, different ages (25 -, 35 -, 45 -, 55 -, ≥65 years old) were compared and analyzed. The number of cases of skeletal fluorosis in Chongqing was calculated according to the 2015 population survey data.Results:A total of 7 768 adults over 25 years old were investigated in 15 villages of 10 townships in 2 counties, and 478 people were diagnosed clinically as skeletal fluorosis, and the clinical detection rate was 6.15%. There were differences in the clinical diagnosis of skeletal fluorosis among different disease areas (χ 2 = 183.23, P < 0.01). There were significant differences among different age groups (χ 2 = 406.73, P < 0.01). But no difference was found among different sex groups (χ 2 = 0.32, P > 0.05). A total of 690 people were diagnosed as skeletal fluorisis by X-ray, the X-ray detection rate was 8.88%, and moderate and severe skeletal fluorosis detection rate was 4.20% (326/7 768). The X-ray diagnosis rates of skeletal fluorosis were different among different disease areas (χ 2 = 46.25, P < 0.01) and different age groups (χ 2 = 384.60, P < 0.01). There was no difference between different sexes groups(χ 2 = 1.77, P > 0.05). According to the different disense in Chongqing, there were about 48 770 cases of skeletal fluorosis diagnosed clinically and 72 630 cases diagnosed by X-ray. Conclusions:The more serious the disease area is, and the older the people's age is, the higher the prevalence of skeletal fluorosis will be. In the future, it is of great important to investigate the prevalence of skeletal fluorosis in coal-burning-borne fluorosis areas in Chongqing.

In this review, development and application of the minimally invasive esophagectomy(MIE) for esophageal cancer are discussed including the types of MIE procedures, short- and long- term outcome after MIE; as well the future of MIE is forecasted. Main procedures of MIE performed currently include esophagectomy via thoracoscopy and laparoscopy and cervical esophagogastrosty, Ivor-Lewis MIE via thoracoscopy and laparoscopy, and hiatal MIE. Ivor-Lewis MIE gradually becomes a standard surgical option for the cancer of distal esophagus or esophagogastric junction while the solution of intrathoracic anastomosis via thoracoscopy has achieved. Several methods of intrathoracic anastomosis are reported such as hand-sewn, circular stapler, side-to-side and triangular anastomosis. MIE could decrease operative blood loss, shorten hospital stay and ICU stay, reduce postoperative especially pulmonary complications, and harvest more lymph nodes compared to open esophagectomy. The long-term survival has been proved similar with that after open esophagectomy for esophageal cancer. MIE has developed rapidly in recent years with some aspects in future prospectively: individual MIE treatment and quality of life, fast track after surgery, and robot-assisted MIE, as well the endoscopic submucosal dissection for esophageal cancer is mentioned.

AJCC Esophageal Cancer Staging System, 8^th edition will be implemented on January 1, 2018. The N staging in 8^th edition of staging system remains following 7^th edition based on the number of metastatic nodes, except the limited revision of the regional lymph node map. N staging revision was reviewed from the simple definition of negative (N0) and positive (N1) lymph node(s) to the positive node number based proposal (7^th edition). The 7^th edition staging system, especially the N staging, were proved with more advantages on distinguishing disease progression and predicting prognosis of the esophageal cancer. On other hand, the disadvantages of 7^th edition N staging are discussed. The refined N staging based on the number of metastatic node station is introduced. The extent and station of metastatic node could better reflect the disease progression and prognosis according to our research. The controversy on N staging of esophagogastric junction cancer is discussed as well. Other reported N staging associated index including lymph node ratio, lymphatic vessel invasion and biomarkers are reviewed and evaluated.