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Fusobacterium nucleatum is an opportunistic pathogenic bacterium that can be enriched in colorectal cancer tissues, affecting multiple stages of colorectal cancer development. The two-component system plays an important role in the regulation and expression of genes related to pathogenic resistance and pathogenicity. In this paper, we focused on the CarRS two-component system of F. nucleatum, and the histidine kinase protein CarS was recombinantly expressed and characterized. Several online software such as SMART, CCTOP and AlphaFold2 were used to predict the secondary and tertiary structure of the CarS protein. The results showed that CarS is a membrane protein with two transmembrane helices and contains 9 α-helices and 12 β-folds. CarS protein is composed of two domains, one is the N-terminal transmembrane domain (amino acids 1-170), the other is the C-terminal intracellular domain. The latter is composed of a signal receiving domain (histidine kinases, adenylyl cyclases, methyl-accepting proteins, prokaryotic signaling proteins, HAMP), a phosphate receptor domain (histidine kinase domain, HisKA), and a histidine kinase catalytic domain (histidine kinase-like ATPase catalytic domain, HATPase_c). Since the full-length CarS protein could not be expressed in host cells, a fusion expression vector pET-28a(+)-MBP-TEV-CarScyto was constructed based on the characteristics of secondary and tertiary structures, and overexpressed in Escherichia coli BL21-Codonplus(DE3)RIL. CarScyto-MBP protein was purified by affinity chromatography, ion-exchange chromatography, and gel filtration chromatography with a final concentration of 20 mg/ml. CarScyto-MBP protein showed both protein kinase and phosphotransferase activities, and the MBP tag had no effect on the function of CarScyto protein. The above results provide a basis for in-depth analysis of the biological function of the CarRS two-component system in F. nucleatum.
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Humans , Histidine Kinase/metabolism , Fusobacterium nucleatum/metabolism , Automobiles , Protein Kinases/genetics , Escherichia coli/metabolism , Colorectal NeoplasmsABSTRACT
Objective To establish an animal model of sparganosis mansoni through oral administration of Cyclops infected with procercoids. Methods Domestic cats were infected with Sparganum mansoni under laboratory conditions, and fresh cat stool samples were collected, washed in dechlorinated water, and filtered. Spirometra mansoni eggs were collected and prepared into suspensions. Twenty C57BL/6j mice were randomly divided into the experimental group (n = 15) and the control group (n = 5). Wild Cyclops were infected with Spirometra mansoni coracidia to allow 3 to 5 procercoids in each Cyclop. Then, each mouse in the experimental group was given 15 Cyclops infected with procercoids by gavage, while mice in the control group were orally administered with the same volume of dechlorinated water. All mice were sacrificed after 5 months, and dissected, and suspicious Sparganum mansoni worms were collected. The serum specific IgG antibody against Sparganum mansoni was measured in mice using enzyme-linked immunosorbent assay (ELISA). Genomic DNA was isolated from suspicious Sparganum mansoni worms, and the specific Sparganum mansoni cytochrome oxidase I (COI) gene was amplified using PCR assay. Results Among the 15 mice in the experimental group, six were positive for the serum specific IgG antibody against Sparganum mansoni, and milky white worms were found and collected from the subcutaneous regions of 4 out of 6 mice. Only one worm was detected in each mouse, and the worm morphology was similar to Sparganum mansoni. Capillary electrophoresis of the PCR amplification products of COI gene presented a specific band with 151 bp in size, and sequencing analysis revealed 100% homology with Sparganum mansoni. Conclusions A mouse model of sparganosis mansoni is successfully created through oral administration of Cyclops infected with Spirometra mansoni procercoids.
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Objective To investigate the prevalence of Spirometra mansoni infections in hosts in Jiangsu Province, so as to provide the scientific basis for the management of sparganosis mansoni. Methods From 2018 to 2019, nine counties (cities, districts) were randomly selected from Jiangsu Province as the survey sites, and 100 healthy individuals were randomly selected to perform the serological test of S. mansoni infections and the detection of S. mansoni eggs. The procercoids were detected in the intermediate host Cyclops, and the S. mansoni eggs were identified in the stool samples of the definitive hosts cats and dogs. Results The prevalence of S. mansoni human infections was 0 (0/900) in the 9 survey sites of Jiangsu Province, and the sero-prevalence of the specific IgG antibody against S. mansoni was 1.22% (11/900). The positive rate of procercoids was 0.33% (3/900) in Cyclops. In addition, the S. mansoni egg-positive rate was 1.48% (2/135) in cats and dogs. Conclusions Sparganosis mansoni is prevalent in Jiangsu Province. Health education pertaining to the damages of sparganosis mansoni and the route of S. mansoni infections should be improved.
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OBJECTIVE: To investigate the effects of nuclear transcription factor-κB(NF-κB)/amide adenine dinucleotide phosphate oxidase 1(NOX1) signaling pathway in tumor necrosis factor-α(TNF-α) induced apoptosis of A549 cells. METHODS: i) A549 cells were stimulated with TNF-α at the concentrations of 0.00, 0.25, 0.50, and 1.00 nmol/L. CCK-8 assay was used to detect the cell viability to screen the optimal stimulating concentration of TNF-α. ii) A549 cells at logarithmic growth stage were randomly divided into four groups, the control group, the TNF-α group, the BAY11-7082(NF-κB inhibitor) group and the TNF-α+BAY11-7082 group. The cells in the control group were not treated. The TNF-α and BAY11-7082 groups were stimulated with 0.50 nmol/L TNF-α and 5 μmol/L BAY11-7082, respectively. The TNF-α+BAY11-7082 group was stimulated by both TNF-α and BAY11-7082. After 24 hours of culture, the cell survival rate was detected by CCK-8 assay. Flow cytometry was used to detect cell apoptotic rate, and Western blotting was used to detect the relative expression of NF-κB(p65) and NOX1 proteins. RESULTS: i) When A549 cells were stimulated with TNF-α at the concentration of 0.50 nmol/L, the cell proliferative activity was reduced and the cell apoptosis was promoted. This concentration was selected as the stimulation dose of TNF-α in subsequent experiments. ii) The survival rate of A549 cells in the TNF-α group decreased(P<0.05), the apoptotic rate and the protein expressions of NF-κB(p65) and NOX1 increased in TNF-α group(all P<0.05) compared with the control group. In BAY11-7082 group, the survival rate and the relative expression of NF-κB(p65) and NOX1 of A549 cells were decreased(all P<0.05), and the apoptotic rate of A549 cells was increased(P<0.05) compared with the control group. A549 cells in TNF-α+BAY11-7082 group changed from a long spindle shape to an irregular one. The cell survival rate increased(P<0.05), the apoptotic rate and the relative expression of NF-κB(p65) and NOX1 decreased(all P<0.05) compared with the TNF-α group. CONCLUSION: NF-κB/NOX1 signaling pathway is involved in A549 cells apoptosis induced by TNF-α.
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Objective:To explore the effects of bilateral arm transcutaneous electrical acupoint stimulation (TEAS) based on mirror therapy (MT) on upper limb function of subacute stroke hemiplegic patients. Methods:From September, 2017 to October, 2019, 48 subacute stroke hemiplegic patients were randomly divided into control group (n = 24) and experimental group (n = 24). All the patients accepted routine rehabilitation and MT, while the experimental group received bilateral arm TEAS and the control group received sham TEAS, for four weeks. They were assessed with Fugl-Meyer Assessment-Upper Extremity (FMA-UE), Action Research Arm Test (ARAT), Wolf Motor Function Test (WMFT) and modified Barthel Index (MBI) before and after treatment. Results:All the scores of FMA-UE, ARAT, WMFT and MBI improved in both groups after treatment (|t| > 11.870, P < 0.001), and improved more in the experimental group than in the control group (|t| > 2.678, P < 0.05). Conclusion:Bilateral arm TEAS based on MT can promote the upper limb function of subacute stroke hemiplegic patients.
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Objective:To investigate the difference of vitamin A and E levels in children with different respiratory diseases at different ages.Methods:A total of 671 children in Hunan Children's Hospital from July 2017 to October 2019 were selected as the disease group, including 197 cases of pneumonia, 152 cases of recurrent respiratory tract infection, 91 cases of asthma, 88 cases of cough variant asthma and 143 cases of Mycoplasma pneumoniae pneumonia; At the same time, 245 healthy children were selected as the normal group. The serum vitamin A and vitamin E levels of the two groups were detected by high performance liquid chromatography (HPLC).Results:⑴ The vitamin A level [(0.31±0.09)mg/L] of the disease group was lower than the normal group [(0.35±0.25)mg/L], and the vitamin E level [(8.92±2.57)mg/L] was lower than the normal group [(9.62±2.79)mg/L], with statistically significant difference ( P<0.05); ⑵ The level of vitamin A in the disease group at the age of >1-3 years [(0.32±0.09)mg/L] was lower than that in the normal group of the same age group [(0.35±0.08)mg/L]; the level of vitamin A in the disease group at the age of >3-6 years old [(0.30±0.08)mg/L] was lower than that of the same age group [(0.32±0.07)mg/L], with statistically significant difference ( P<0.05); ⑶ The vitamin E level of the disease group at >1-3 years old [(9.23±2.56)mg/L], >3-6 [(8.02±1.86)mg/L] and >6-14 years old [(8.02±1.82)mg/L] were lower than that of the same age normal group [(9.76±2.81)mg/L, (9.67±2.87)mg/L, (9.19±2.58)mg/L], with statistically significant difference ( P<0.05); ⑷ There were significant differences in vitamin A levels among different age in disease group ( P<0.05). Among them, the children with high risk of subclinical deficiency accounted for the largest proportion (45.78%) in the 6-month-1-year-old group, and the proportion of children with normal vitamin A levels in other age groups was the largest; ⑸ There are significant differences in vitamin E levels in different age groups in the disease group ( P<0.05), the levels in the normal range accounts for the largest proportion of all ages; ⑹ The levels of vitamin A and vitamin E in mycoplasma pneumoniae infection group were increased compared with in recurrent respiratory infection group , asthma group, and cough variant asthma group, and the difference was statistically significant ( P<0.05). Compared with the pneumonia group, the level of vitamin E increased in the recurrent respiratory infection group, and the difference was statistically significant ( P<0.05); The vitamin E levels in the cough variant asthma group were reduced compared with the repeated respiratory infection group, asthma group and pneumonia group ( P<0.05). Conclusions:The Vitamin A and E levels of children suffering from respiratory diseases are lower than those of normal children. The Vitamin A and E levels of different respiratory diseases and different age groups are different. Vitamin A and E supplementation may be significantly targeted according to different ages and different respiratory diseases in clinical practice.
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Background@#Although a number of technical problems and donor safety issues associated with living donor liver transplantation (LDLT) have been resolved, some initial clinical studies showed an increased risk of hepatocellular carcinoma (HCC) recurrence in LDLT. This meta-analysis was conducted to assess differences in tumor recurrence between LDLT and deceased donor liver transplantation (DDLT).@*Methods@#After systematic retrievals of studies about LDLT and DDLT for HCC, articles were selected with a rationale of emphasizing inter-group comparability. Results from multivariate analyses were combined and discussed together with univariate analyses. In subgroup analysis, the impact of organ allocation policy was taken into consideration.@*Results@#Seven articles were included in the meta-analysis. Overall, a salient result that emerged from the seven studies was a significant increased risk of HCC recurrence in the LDLT group than in the DDLT group (P = 0.01). The most significant increase in hazard ratio was found in studies where organs tended to be allocated to non-tumor patients.@*Conclusions@#An increased risk for HCC recurrence in LDLT as compared with DDLT patients was found. The relatively shorter preoperative observation windows in LDLT may lead to fewer cases of HCC with invasive features being screened out, which may provide a possible explanation for the high rates of HCC recurrence.
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Novel antineoplastic drugs have significantly prolonged the survival time of cancer patients. Meanwhile, nephrotoxicity of antineoplastic drugs and its adverse effects on the prognosis of cancer patients have received increasing attention. Conventional chemotherapy causes kidney injury mainly through direct renal toxicity, while new anti-tumor drugs can induce a number of kidney damage, including acute renal tubular injury, thrombotic microangiopathy, interstitial nephritis, and glomerular diseases through multiple mechanisms. Clinicians must be knowledgeable in the renal toxicity of antineoplastic drugs to minimize the nephrotoxicity of the drugs and diagnose early, especially in patients with underlying kidney disease. This article focuses on the risk factors, clinical and histological patterns, pathogenesis, prevention and treatment of renal injury associated with the antineoplastic drugs, especially novel targeted antineoplastic drugs.
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Gastrointestinal stromal tumors (GISTs) are the most common mesenchymal tumors. Surgery remains to be the primary treatment for patients with localized GISTs, but there are still many patients suffering from tumor metastasis and recurrence after surgery. Imatinib adjuvant therapy plays an important role in the prevention and treatment of tumor metastasis and recurrence, but there are still many controversies in terms of dosage and time of administration. For initial unresectable GISTs with large volume, neoadjuvant therapy may considered to be an option. Sunitinib and regorafenib have played an important role in the second and the third line treatments. BLU-285 has brought hope to patients with mutation of PDGFRA D842. The emerging immunotherapy is still in the exploration stage of gastrointestinal stromal tumors in recent years. This article reviews the research progress of surgical treatment, neoadjuvant therapy, postoperative adjuvant therapy and other treatments for GISTs.
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OBJECTIVE: To study a potential relationship between preterm labor and lymphocyte to monocyte ratio(LMR).METHODS: This retrospective cohort study was conducted in Second Affiliated Hospital of Chongqing Medical University from August 2016 to November 2017. Totally 100 pregnant women who delivered between 28 th to 37 th gestational week were included as the study group,and 116 pregnant women who delivered after 37 th gestational week were as the control group. The data of routine blood test from 11 th to 13 th gestational week and 28 th to 30 th gestational week were observed and compared.RESULTS: The LMR value of preterm women was significantly higher than that of those who delivered at term(early pregnancy 4.90±1.40 vs. 4.31±1.30,P<0.01;middle and advanced stage of pregnancy 3.54±0.93 vs. 2.95±0.64,P<0.01). A negative correlation was observed between the level of LMR and the gestational weeks of termination of pregnancy(r=-0.350,P<0.01).CONCLUSION: The value of LMR in pregnancy is associated with the occurrence of preterm labor. Therefore,it is necessary to make further study.
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BACKGROUND@#Although a number of technical problems and donor safety issues associated with living donor liver transplantation (LDLT) have been resolved, some initial clinical studies showed an increased risk of hepatocellular carcinoma (HCC) recurrence in LDLT. This meta-analysis was conducted to assess differences in tumor recurrence between LDLT and deceased donor liver transplantation (DDLT).@*METHODS@#After systematic retrievals of studies about LDLT and DDLT for HCC, articles were selected with a rationale of emphasizing inter-group comparability. Results from multivariate analyses were combined and discussed together with univariate analyses. In subgroup analysis, the impact of organ allocation policy was taken into consideration.@*RESULTS@#Seven articles were included in the meta-analysis. Overall, a salient result that emerged from the seven studies was a significant increased risk of HCC recurrence in the LDLT group than in the DDLT group (P = 0.01). The most significant increase in hazard ratio was found in studies where organs tended to be allocated to non-tumor patients.@*CONCLUSIONS@#An increased risk for HCC recurrence in LDLT as compared with DDLT patients was found. The relatively shorter preoperative observation windows in LDLT may lead to fewer cases of HCC with invasive features being screened out, which may provide a possible explanation for the high rates of HCC recurrence.
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This article introduces the process and principles of variety selection for medical device supervision and inspection, and it analyzes the reason and consideration of the variety selection data of national medical device supervision and inspection from 2017 to 2019, it also put forward a collection of selected varieties for reference in the future by sorting out the classified catalogue of medical devices and diagnostic reagents.
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Equipment SafetyABSTRACT
This paper reviews the quality and safety risk points of medical devices found in the sampling inspection of national medical device supervision in recent years. These risk points are summarized into quality management system, product technical requirements, national standards and industry standards, etc. Several specific risk scenarios are further summarized. Based on the above efforts, a more comprehensive risk point system is constructed. Then it is illustrated with typical examples. In view of the existing problems, the corresponding suggestions are put forward to the production enterprises and supervision departments respectively.
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Equipment Safety , Equipment and Supplies , Industry , Reference StandardsABSTRACT
Exosomes are a group of membraneous vesicles generated and released by multi-vesicular bodies or cell membranes in a variety of cell types. Acting as important messages between cells, they participate in almost every physiological and pathological process of living organisms. Exosomes contain specific proteins, mRNA, miRNAs, etc. and mediate intercellular communications, signal transductions and gene expressions effectively. Exosomes are involved in the formation of hepatic fibrosis, which is the typical liver pathological change in the progression of schistosomiasis and is caused by the liver repair and (or) regeneration involving inflammation stimulated by exosomes, activated hepatic stellate cells and other related pathways in reaction to the parasite infection. Exosomes could serve as new markers for schistosomiasis hepatic fibrosis diagnosis and potential targets for its treatment. This paper briefly reviews the latest development of studies on the regulatory roles of exosomes in schistosomiasis hepatic fibrosis, so as to provide ideas for searching new treatment targets of the disease.
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Exosomes are a group of membraneous vesicles generated and released by multi-vesicular bodies or cell membranes in a variety of cell types. Acting as important messages between cells, they participate in almost every physiological and pathological process of living organisms. Exosomes contain specific proteins, mRNA, miRNAs, etc. and mediate intercellular communications, signal transductions and gene expressions effectively. Exosomes are involved in the formation of hepatic fibrosis, which is the typical liver pathological change in the progression of schistosomiasis and is caused by the liver repair and (or) regeneration involving inflammation stimulated by exosomes, activated hepatic stellate cells and other related pathways in reaction to the parasite infection. Exosomes could serve as new markers for schistosomiasis hepatic fibrosis diagnosis and potential targets for its treatment. This paper briefly reviews the latest development of studies on the regulatory roles of exosomes in schistosomiasis hepatic fibrosis, so as to provide ideas for searching new treatment targets of the disease.
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The types and the reasons of changing the original test result after retesting in national medical device sampling and testing from 2013 to 2016, are summarized and analyzed. Firstly, collecting data of "standards not complied" and retesting. Then, giving specific examples when summarizing five types of changing the original test result after retesting. Meanwhile, analyzing the relevant reasons, discovering the deep problems. Finally, giving suggestions for the above problems.
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Equipment and Supplies , Reference Standards , Quality ControlABSTRACT
Objective: To investigate the effects of dexamethasone on transforming growth factor [TGF]-beta1/Smads signaling pathway in benign biliary stricture [BBS] fibroblasts
Study Design: An experimental study
Place and Duration of Study: Guizhou Medical University, Guiyang, Guizhou, China, from January to August 2016
Methodology: Fibroblasts derived from rabbit BBS model were cultured and identified, then treated by different concentration of dexamethasone [0.02, 0.1 and 0.5 mg/ml]. Dexamethasone-treated cells and non-treated control groups were incubated respectively for 48 hours. Cell proliferation was assessed using cell counting kit-8. Relative mRNA expression of TGF-beta1, Smad4 and Smad7 were assessed by quantitative RT-PCR. Protein expression of TGF-beta1 and Smad4 were investigated by Western blotting
Results: Treatment with dexamethasone significantly inhibited the proliferation of BBS fibroblasts, significantly attenuated both the mRNA and protein expression of TGF-beta1 and Smad4, and significantly up-regulated the mRNA expression of Smad7 in BBS fibroblasts [all p<0.05, 0.1-0.5 mg/ml], and exhibited in a dose-dependent manner
Conclusion: TGF-beta1/Smads signaling pathway may play an important role in BBS progression; dexamethasone significantly altered the expression of TGF-beta1/Smads signaling pathway and significantly inhibited cell proliferation in rabbit BBS fibroblasts. Therefore, dexamethasone may be a therapeutic option for the prevention of BBS
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Aim To isolate HeLa cell proliferation-inhibitory active fraction from Pegasus laternarius Cuvier and explore its potential apoptosis-inducing mechanism.Methods To obtain the active fraction,the ethanol extract of Pegasus laternarius Cuvier was chromatographed by silica gel and sephadex LH-20 columns;MTT assay was used to evaluate the proliferation-inhibitory ability of active fraction on HeLa cells;AO/EB,PI and Annexin V-FITC/PI fluorescent staining flow cytometry were used to evaluate its apoptosis-inducing ability;the possible mechanism was investigated by analyzing the enzyme activity of caspase-3 and the protein expression of apoptosis-related genes in tumor cells.Results A fraction of C22 with high HeLa proliferation-inhibitory activity was isolated,with a yield of 0.73 ‰ and an IC50 of 36.3 mg · L-1;fraction C22 could increase the proportion of cells in sub-G0/G1 phase,phosphatidylserine eversion and other typical cell apoptosis in a dose-dependent manner;fraction C22 could down-regulate the expression of Bcl-2 and increase the enzyme of caspase-3 in HeLa cells.Conclusions The active fraction C22 from Pegasus laternarius Cuvier can inhibit the proliferation of HeLa cells by inducing apoptosis.The effect of inducing apoptosis may be conducted through mediating the mitochondrial Bcl2/caspase pathway.
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Objective: To investigate the effects of tetramethylpyrazine [TMP] on transforming growth factor-beta1 [TGF- beta1], alpha-smooth muscle actin [alpha-SMA], and neuronal regeneration related protein [P311] in benign biliary stricture fibroblasts of rabbit. Study Design: An experimental study. Place and Duration of Study: Guizhou Medical University, Guiyang, Guizhou, China, from April to December 2015
Methodology: Fibroblasts isolated from rabbits following benign biliary stricture were cultured and treated with different concentrations of TMF [0.08, 0.4. and 2.0 mg/ml]. TMP-treated cells and non-treated control groups were incubated for 48-hours, and proliferation was assessed using the cell counting kit-8 assay. The mRNA expressions of TGF-beta1, alpha-SMA, and P311 were assessed by quantitative RT-PCR. Protein expressions of TGF-beta1 and alpha-SMA were investigated by Western blotting
Results: Treatment with TMP significantly reduced the proliferation of benign biliary stricture fibroblasts, and significantly attenuated both the mRNA and protein expressions of TGF-beta1, alpha-SMA, and P311 [p <0.05] in a dose-dependent manner
Conclusion: TMP significantly reduced the proliferation of benign biliary stricture fibroblasts, and significantly down- regulated the mRNA/protein expression of TGF-beta1, alpha-SMA, and P311. Therefore, TMP may be a therapeutic option for the prevention of benign biliary stricture
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Objective To observe the inhibitive effect of electro-acupuncture (EA)at Zusanli points (ST36)on inflammatory mediators of postoperative intra-abdominal adhesions and study the relationship between EA and cholinergic anti-inflammatory pathway.Methods Forty-eight male Wistar rats were divided into 6 groups (each =8):Group A (control),Group B(abdominal adhesions model),Group C (abdominal adhesions plus EA),Group D(sham acu-point control),Group E (abdominal adhesions plus α-bungarotoxin )and Group F (abdominal adhesions plus EA after α-bungarotoxin).Animal models of abdominal adhesion were produced by Chiang’s path.Bilateral Zusanli points (ST36) and shame acupoints were electro-acupunctured at a constant voltage for 1 hour while rats were awake.The ɑ-BGT(1 μg/kg)was injected into the abdominal cavity after surgery.All the rats were sacrificed on the 3rd day,and the levels of inflammatory mediators (TNF-ɑ,NO and NOS)in tissues were evaluated.Results Three days after surgery,the damaged cecum of abdominal adhesion groups developed obvious edema that did not adhere with other tissues.Compared with sham control,the abdominal adhesion resulted in significant elevation of inflammatory mediators (TNF-ɑ,NO and NOS).EA at Zusanli points obviously lowered the elevated levels of inflammatory mediators (P <0.01 and P <0.05).EA at Zusanli points following the injection of ɑ-BGT showed less anti-inflammatory effect(P <0.01).Conclusion EA at Zusanli points significantly lowers the elevated levels of inflammatory mediators after abdominal adhesion challenge.The activation of cholinergic anti-inflammatory pathway might be one of the mechanisms by which Zusanli points exert anti-inflammatory effects.