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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 compare the effects of hip abductor fatigue on postural stability and neuromuscular control in different gender groups during single-leg side-jump landing. Methods Twenty male and twenty female adults were required to execute single-leg side-jump landing before and after hip abductor fatigue protocol. The center of pressure (COP), ground reaction force (GRF), lower extremity kinematics, joint moment, muscle activity were compared. Results For both males and females after fatigue, the maximum displacements and average velocities of COP decreased, and the peak angles of hip abduction and ankle eversion increased, and the peak torques of ankle inversion increased. At 200 ms pre-landing, the activiation of rectus femoris, biceps femoris, tibialis anterior, peroneus longus were lower in male than that in female. At 200 ms post-landing, the activiation of biceps femoris in male was lower than that in female after fatigue. Conclusions The postural stability in frontal plane decreased after hip abductor fatigue. The frontal plane control of hip joint and ankle joint was weakened, which might increase the risk of joint injury. There are differences in postural strategies for different genders, indicating that gender difference in the mechanism of lower limb joint injury is worth of further investigation.
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Objective To discuss the angiographic manifestations of hemorrhage after pancreaticoduo-denectomy (PD), and to evaluate the clinical curative effect of interventional therapy. Methods The angiographic findings and the therapeutic effect of interventional therapy for hemorrhage after PD in 19 PD patients, who were admitted to Fujian Provincial Hospital, China, during the period from January 2014 to February 2018 to receive DSA examination and interventional therapy, were retrospectively analyzed. Results Among the 19 PD patients, DSA examination showed that extravasation of contrast medium, pseudoaneurysm formation and irregular arterial lumen were observed in 16 patients, the rate of positive signs was 84.2％ (16/19) . All the 16 patients, who had positive DSA findings, received covered-stent implantation or embolization therapy for the responsible artery, except one patient who had hemorrhage from the branch of superior mesenteric artery and the super-selective catheterization for him failed. The technical success rate was 93.8％ (15/16) . Successful hemostasis was achieved in 13 patients (86.7％, 13/15) . Two patients developed recurrent bleeding and were transferred to surgical treatment. Two patients received twice angiography, the resultsshowed that pseudoaneurysm formation of gastroduodenal artery was detected in one patient, which was treated with embolization therapy, and in another patient angiography was normal and this patient was transferred to surgical treatment. Conclusion For postoperative hemorrhage of PD, DSA has diagnostic value, while interventional therapy has therapeutic value. The techniques are minimally-invasive and highlyeffective. Therefore, it is worthy of clinical popularization and application.
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Aim To explore the protective effects of naringin on hypoxic ischemic brain injury in neonatal rats and its mechanism. Methods Ninety-six healthy 7-day neonatal SD rats were randomly divided into sham operation group, hypoxic-ischemic brain damage group (HIBD group),HIBD with low-dose naringenin group(50 mg·kg-1, NG-L) and HIBD with high-dose naringenin group(100 mg·kg-1,NG-H). Neu-rological deficit, HE staining and brain water content were measured 48h after operation. Immunoblotting was used to detect the expressions of NOD2,RIP2 and NF-κB. Enzyme linked immunosorbent assay(ELISA) method was adopted to detect TNF-α and IL-1β ex-pression. Results Compared with HIBD group, the neurological deficit score decreased, the pathological damage was reduced, and the water content of brain tissues markedly decreased by naringenin(50,100 mg ·kg-1) treatment(P<0.05). Western blot revealed the down-regulation of NOD2,RIP2 and NF-κB by na-ringenin (50,100mg·kg-1) treatment (P<0.05). The content of TNF-α and IL-1β in brain tissues was lower than that of HIBD group (P <0.05). Conclu-sion Naringenin is likely to exert a protective role in neonatal rats of hypoxic ischemic brain injury perhaps through decreasing the expression of NOD2, RIP2 and NF-κB,and reducing the secretion of TNF-α and IL-1β.
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Objective To investigate the feasibility of sequential intensity-modulated radiotherapy (sIMRT)and simultaneously integrated boost intensity-modulated radiotherapy(SIB-IMRT)in the radiotherapy of brain metastasis,the dosimetric difference of target volumes and organs at risk(OARs). Methods Twenty pa-tients diagnosed as brain metastasis were randomly selected,with SIB-IMRT and sIMRT programs developed for each patient. Dosimetric differences between target areas and OARs were compared between the two radiotherapy protocols. Results Compared with sIMRT,SIB-IMRT had no significant difference in the average irradiation dose of the brainstem[(42.69 ± 2.18)Gy vs.(41.98 ± 0.96)Gy]and homogeneity index(HI)(1.46 ± 0.04 vs.1.42 ± 0.13)of P-CTV(P > 0.05). However,SIB-IMRT plan achieved higher than sIMRT in the conformation index (CI)(0.68 ± 0.05 vs. 0.44 ± 0.04)and HI(1.03 ± 0.01 vs. 1.06 ± 0.01)of P-GTV. Meanwhile,both maximum exposure dose of OARs and CI of P-CTV(0.68 ± 0.05 vs.0.44 ± 0.04)of SIB-IMRT were significant in comparison with sIMRT(P<0.05).Conclusions Both radiotherapies can meet target coverage and dose requirements.Com-pared to sIMRT technique,SIB-IMRT technique can decrease effectively the exposure dose of surrounding organs, and can give the tumor target more uniform physical dose conformation.
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Aim To study the therapeutic effect of CpG-ODN, an agonist of Toll-like receptor 9 (TLR9), on hypoxic/ischemic encephapathy in neonatal rats and investigate the mechanisms.Methods Fifty healthy 7-day-old neonatal Wistar rats (in either gender, weighing 12~17g) were randomly divided into sham operation group, HIBD group, and CpG-ODN low group(0.35 mL·kg-1), CpG-ODN middle group(1.40 mL·kg-1), CpG-ODN high group(5.60 mL·kg-1).The neurological function was scored after 48h operation;ten rats of each group was executed respectively and brains tissue was taken;HE staining was used to observe the brain pathological changes.Western blot assay was used to detect the expressions of TLR9 and phosphor-p38 mitogen-activated protein kinases(p-p38 MAPK), and enzyme linked immunosorbent assay (ELISA) method was adopted to detect TNF-α expression.Results The CpG-ODN low, middle group were improved in impairment significantly compared with the HIBD group, and the brain pathological change was lessened, while the CpG-ODN high group was impaired significantly compared with the HIBD group (P<0.05), and brain pathological change was sharpened.Western blot showed the up-regulation in TLR9 and p-p38 MAPK and a significant increase of the expression of TNF-α in the brain tissue in CpG-ODN group with statistical difference in HIBD group and sham operation group(P<0.05).Conclusions The neuro-behavioral score and nervous system function can be improved and the hypoxic/ischemic brain damage can be reduced in neonatal rats in the CpG-ODN low, middle group.The protective mechanisms may be suitably via activating p38 MAPK signaling pathway to promote p38 MAPK phosphory1ation and up-regulation of the expression of TNF-α in the brain tissue of rats.
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Objective To evaluate the therapeutic efficacy of percutaneous transhepatic portal vein balloon angioplasty in treating cavernous transformation of portal vein following operation of congenital choledochal cyst. Methods From 2012 to 2014, a total of 6 patients with cavernous transformation of portal vein which occurred after the operation of congenital choledochal cyst were encountered at authors’ hospital. The clinical data were retrospectively analyzed. Before treatment, all patients presented symptoms of different degrees of hematemesis. Percutaneous transhepatic portal vein balloon angioplasty was carried out in all patients, and embolization of gastric coronary vein with coils was employed if angiography showed that coronary vein of stomach was pronouncedly dilated. The clinical manifestations, the imaging materials and the complications were analyzed. All the patients were followed up for 3 - 31 months. Results Of the 6 patients, portal vein main stem occlusion was found in 5 and severe localized stenosis was seen in one. Cavernous transformation of portal vein was revealed in all the 6 patients. Percutaneous transhepatic portal vein balloon angioplasty was successfully accomplished in 5 patients and failed in one patient. Embolization of gastric coronary vein with coils was performed in two patients. After the treatment, no treatment-related severe complications occurred in all patients. The follow-up period ranged from 3 to 31 months. During the follow-up period portal vein maintained patent in 5 patients. No recurrent hematemesis occurred in all patients. Conclusion For the treatment of cavernous transformation of portal vein occurring after the operation of congenital choledochal cyst, percutaneous transhepatic portal vein balloon angioplasty is a safe, effective and minimally - invasive therapeutic means.
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<p><b>OBJECTIVE</b>To retrospectively analyze the safety and efficacy of mechanical thrombectomy combined with pharmacologic thrombolysis to treat non-acute and symptomatic portal vein thrombosis (PVT) using an intrahepatic portosystemic shunt (IPS) assisted by percutaneous transhepatic approach.</p><p><b>METHODS</b>From April 2006 to May 2012, 18 patients with non-acute and symptomatic PVT were treated with balloon dilation, sheath-directed thrombus aspiration and continuous infusion of urokinase using the IPS assisted by percutaneous transhepatic approach. The significance of differences in the portosystemic gradient measured before and after therapy was assessed by paired samples t-test, and survival analysis was made by the Kaplan-Meier method.</p><p><b>RESULTS</b>IPS was successfully created in all patients. The mean duration of the thrombolytic therapy was 65.3 +/- 29.5 h, and the mean concentration of urokinase used for the thrombolysis was 2324000 +/- 945000 U. Comparison of the mean portosystemic gradients showed a significant improvement in response to the therapy (before: 33.8 +/- 4.9 mm Hg vs. after: 15.4 +/- 2.1 mm Hg; P less than 0.001). The overall rate of clinical improvement was 94.4%. One patient died on day 2 post-therapy and another two patients experienced mild hepatic encephalopathy or right hemothorax, respectively, on day 5 post-therapy, with conservative medical management achieving complete recovery for both. The mean follow-up time was 18.6 +/- 17.5 months, during which only one patient died and five others experienced shunt dysfunction; all remaining patients showed maintenance of shunt patency without symptoms of recurrence.</p><p><b>CONCLUSION</b>Mechanical thrombectomy combined with pharmacologic thrombolysis via the IPS assisted by percutaneous transhepatic approach is a safe and effective therapeutic option for patients with non-acute and symptomatic PVT.</p>
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Adult , Aged , Female , Humans , Male , Middle Aged , Portal Vein , Portasystemic Shunt, Surgical , Methods , Retrospective Studies , Thrombolytic Therapy , Venous Thrombosis , TherapeuticsABSTRACT
<p><b>OBJECTIVE</b>To compare the efficacies of portal vein stenting and transcatheter arterial chemoembolization (TACE) combined therapy performed with or without endovascular implantation of iodine-125 (125I) seeds strand in patients with hepatocellular carcinoma (HCC) and main portal vein tumor thrombus (MPVTT).</p><p><b>METHODS</b>One-hundred-and-six patients with HCC complicated by MPVTT who were treated with portal vein stents and TACE, either with (Group A, n=56) or without (Group B, n=50) endovascular implantation of 125I seeds strand, between July 2005 and April 2011, were retrospectively analyzed. Overall survival, stent patency, and procedure-related adverse events were compared between the two groups.</p><p><b>RESULTS</b>The technical success rate was 100% for placement of 125I seeds strands and stents in the obstructed main portal vein. No serious procedure-related adverse events were recorded. Group A had significantly higher median survival (335 days vs. group B: 146 days; P=0.001, hazard ratio (HR)=2.244). Additionally, group A had significantly higher median stent patency (400 days vs. group B: 190 days; P=0.005, HR=2.479).</p><p><b>CONCLUSION</b>The combination therapeutic strategy of portal vein stenting and TACE with endovascular implantation of 125I seeds strands improves the survival of HCC patients with MPVTT complication.</p>
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Female , Humans , Male , Middle Aged , Carcinoma, Hepatocellular , Therapeutics , Chemoembolization, Therapeutic , Combined Modality Therapy , Iodine Radioisotopes , Therapeutic Uses , Liver Neoplasms , Therapeutics , Neoplastic Cells, Circulating , Portal Vein , General Surgery , Retrospective Studies , Stents , Treatment Outcome , Venous Thrombosis , TherapeuticsABSTRACT
ObjectiveTo establish an animal model of implanted inferior vena cava tumor thrombus (IVCTT)and examine its growth with MDCT and 3D-MPR. MethodsTumor cell line VX2 was inoculated subcutaneously into rabbit to develop the primary tumor, which was then cut into small strips. Purse-string suture was performed on the anterior wall of IVC after the laparotomy in eighteen New Zealand white rabbits.The tumor strip was injected into IVC through the purse and suspensory fixed on the inner wall of IVC. The general conditions,body weight,and the survival time were monitored after operations.MDCT examinations were performed with plain scan,arterial phase,portal phase and venous phase enhancement every week for all animals and 3D-MPR were acquired.The volumes of IVCTT were calculated.IVC,IVCTT and metastasis were examined with gross and histological pathology. ResultsThe IVCTT was confirmed by MDCT and 3D-MPR images.Collateral varicose veins caused by IVC obstruction and metastasis were also shown in images.IVCTT and metastasis were confirmed by pathological method. The success rate of IVCTT was 100 ％. The mean survival time of operated animals was(49.5±4.4)days. ConclusionsInjecting and suspensory fixing VX2 tumor strip into IVC is a reliable method to establish the IVCTT animal model. MDCT and 3D-MPR are valuable methods to monitor the growth and metastasis of IVCTT in animal models. The model of implanted IVCTT of rabbits provides a useful tool for the research of treatment of IVCTT.
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<p><b>OBJECTIVE</b>To study the effects of rhizoma sparganii and radices zedoariae on hepatic fibrosis.</p><p><b>METHOD</b>The rat immunohepatic fibrosis model was made by intraperitoneal injection of porcine serum and treated with rhizoma sparganii and radices zedoariae. The ALT, GGT, TP, ALb, A/G, IVC, LN, HA and the pathological change of the liver were observed.</p><p><b>RESULT</b>Rhizoma sparganii and radices zedoariae could increase TP, ALb, A/G, decrease ALT, GGT, IVC, LN, HA and improve the pathological change.</p><p><b>CONCLUSION</b>Rhizoma sparganii and radices zedoariae can protect hepatic cells, alleviate degeneration and necrosis, recover structure and function, and reduce the proliferation of fibrous tissue.</p>