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This study mainly explores the role of myeloid differentiation primary response protein 88 (MyD88) in tumorigenesis and development, to identify active compounds targeting MyD88. CRISPR/Cas9 system and xenograft tumor model were used to detect the effect of MyD88 deletion on tumor growth, and the experimental animal ethics review number was PZSHUTCM200828006. Microscale thermophoresis technology (MST) was used to identify compounds directly bind to MyD88 and further detect the impact of candidate small molecules on cell proliferation. Results showed that depletion of MyD88 significantly inhibited xenograft tumor growth of colon cancer, pancreatic cancer and skin cancer and the activity of NF-κB signaling pathway. MST showed that nordihydroguaiaretic acid (NDGA) bound to MyD88, with the binding dissociation constant Kd of 14.61 µmol·L-1. NDGA inhibited NF-κB reporting system activation and phosphorylation of p65, the key factor in NF-κB signal pathway. In addition, the results of colony formation assay showed that NDGA suppressed the proliferation of tumor cells. The above results show that, MyD88 is a potential therapeutic target for colon cancer, pancreatic cancer and skin cancer, NDGA directly binds to MyD88 and inhibits the activity of NF-κB signaling pathway, as well as inhibits the proliferation of pancreatic cancer, skin cancer and colon cancer cells.
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Huachansu is a traditional Chinese medicine widely used in the clinic for cancer therapy, while the underlying mechanism is not fully clarified. This study was to investigate the targets and mechanisms of cinobufagin (CBG), an active component of Huachansu, in terms of blocking mitosis of cancer cells. Propidium iodide (PI) DNA staining was used to analyze the effect of CBG on cell cycle. The effect of CBG on mitosis of cancer cells was examined by α-tubulin and pericentrin staining after synchronization by a double thymidine block. Tubulin turbidity, tubulin polymerization and α-tubulin immunofluorescence assays were used to evaluate the effect of CBG on microtubule polymerization. CRISPR/Cas9 gene-editing technology was used to knockout microtubule-severing protein Katanin regulatory subunit B1 (KATNB1) in HCT116 cells, and the inhibitory effect of CBG on wild-type cells and knockout cells was measured by CCK-8. The engagement of CBG with KATNB1 was measured by CETSA and DARTS assays. The effect of CBG on KATNB1 protein and mRNA level was examined by Western blot and real-time PCR, respectively. Our data showed that CBG arrested HCT116 cell cycle at the G2/M phase, disrupted mitosis and induced centriole overduplication. CBG significantly inhibited tubulin polymerization in vitro and in vivo. The cytotoxicity of CBG inhibition on HCT116 was significantly attenuated upon KATNB1 depletion. Moreover, CBG bound to KATNB1 and decreased its protein level, while mutated KATNB1 weakened this effect. In conclusion, CBG inhibited microtubule polymerization via targeting KATNB1, thereby disrupting mitosis in cancer cells.
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Overdenture as a treatment modality for both partially and fully edentulous patients is costeffective and less expensive.The purpose of the present study was to examine the newly fabricated attachments by comparing them with conventional O-ring attachment in vitro in terms of retention force and cyclic aging resistance.A total of 150 samples were prepared and divided into five groups according to the materials used (O-ring attachment,Deflex M10 XR,Deflex Classic SR,Deflex Acrilato FD,and flexible acrylic resin).The retention force of different attachments was measured by a mini dental implant after three subsequent aging (0,63,and 126) cycles in the circumstances similar to the oral environment.The gap space between the head of the implant and the inner surface of the attachments was detected.Two-way analysis of variance (ANOVA) analysis with multiple comparisons test was applied for statistical analysis.The results showed that Deflex M10 XR had the highest retention force and the lowest gap space after cyclic aging;in addition,by comparing the relative force reduction,the lowest values were obtained in the O-ring attachment and the highest values in the flexible acrylic resin attachment.The retention force measured after cyclic aging for the Deflex M10 XR attachment was greatly improved when compared with the O-ring attachment and other types of attachment materials;in addition,the Deflex M10 XR attachment exhibited the minimum gap space between the inner surface and the mini dental implant head.In conclusion,Deflex M10 XR has the ability to withstand weathering conditions and retains its durable and retentive properties after aging when compared with other attachments.
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Objective To explore the clinical valuation and feasibility of transumbilical abdominal wall within the three-channel single-incision laparoscopic combined cholecystectomy and appendectomy.Methods The clinical data of 40 patients who underwent transumbilical abdominal wall within the three-channel single-incision laparoscopic combined cholecystectomy and appendectomy from January 2011 to March 2013 were retrospectively analyzed.Results Forty patients were successfully completed surgery,no transfer laparotomy and death cases,surgery time was 60-80 min,average 70 min,without postoperative incision infection,wound bleeding,bile duct injury,appendix stump fistula and abdominal residual abscess,neither postoperatively used analgesics,1 month after without incision scar,cosmetic effect was remarkable.Half a year's follow-up,the excellent operation evaluation.Conclusions The performer should have accomplish laparoscopic operation skills,and master the indications and the principle.The method of transumbilical abdominal wall within the three-channel single-incision laparoscopic combined cholecystectomy and appedectomy is safe,feasible and valid.The postoperative wound is more minimally invasive and aesthetic effect is obvious.
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Previously,the choice of prosthetic implant-retained overdentures has depended on data from previous studies about the retention-fatigue strength of the attachment system selected.Little or no data have been available on the correlation between the attachment system selected and the overdenture support configuration.The purpose of the present study was to evaluate the retention force and fatigue resistance of three attachment systems and four support designs of overdenture prosthesis.Four lower edentulous acrylic models were prepared and eight combinations of attachments groups were investigated in the study.These included:O-Rings with mini-dental implants (MDIs),Dalbo elliptic with Dalbo Rotex and fabricated flexible acrylic attachments with both MDI and Dalbo Rotex.The study was divided into four test groups:groups A and B,controls,and groups C and D,experimental groups.Control group A contained three overdenture supports:two free standing MDIs in the canine region and at the midline,and one simulated tooth root with Dalbo Rotex screwed in.Control group B contained four overdenture support foundations:two free standing MDIs in the right canine region and the first premolar region,and two simulated tooth roots with Dalbo Rotex screwed in at the same MDI position,but on the left side of the model.Experimental group C contained three overdenture support foundations:two free standing MDIs in the canine region and at the midline,and one simulated tooth root with MDI screwed in.Experimental group D contained four overdenture support foundations:two free standing MDIs in the right canine region and the first premolar region,and two simulated tooth roots with MDIs screwed in at the same MDI position,but on the left side of the model.Each group was further divided into two subgroups according to attachment type used.Five samples were prepared for each group.Retention force (N) values were recorded initially (0 cycles) and after 360,720,1440 and 2880 insertion and removal cycles.During the tensile test a cross-head speed of 10 mm/min was applied.Values of absolute force (AF) and relative force (RF) were statistically analyzed by two-way ANOVA and multiple comparison Tukey's tests between groups and cycles periods.The results of fatigue tests showed a 50％ reduction in retention force in the subgroups with flexible attachments.A triangular design of overdenture support foundations with O-Ring attachments revealed the lowest value of AF and a relatively high reduction in RF.The four overdenture support designs with flexible acrylic attachments improved the retention force and reduced the fatigue retention.Furthermore,the results of the investigation demonstrate that flexible acrylic attachments for both teeth and implant-supported overdentures offer a wide range of retention forces.
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The purpose of this study was to evaluate the effect of bone marrow mesenchymal stem cells(MSCs)transfected with the basic fibroblast growth factor(bFGF)-expressing recombinant adeno-associated virus vector(rAAV2-bFGF),on early angiogenesis of calvarial defects in rats.The MSCs were cultured and transfected with rAAV2-bFGF after differential adherence isolation.The transfection efficiency was detected by RT-PCR and Western blotting.The transfected MSCs were compounded with poly-DL-lactide/hydroxyapatite(PDLLA/HA)in vitro.The cranial defect models in36 malc SD rats were created.Nothing(group A),PDLLA/HA alone(group B),PDLLA/HA combined with MSCs(group C),and PDLLA/HA combined with rAAV2-bFGF transfected MSCs(group D)were implanted in rat calvarial defects.The specimens were harvested for hematoxylin-eosin staining on the day 1,3 and 7 after implantation.Factor Ⅷ immunohistochemical staining and histomorphometric analysis were carried out to evaluate neovascularization around the implantation.The results indicated that MSCs could indeed be successfully transfected with the rAAV2-bFGF vector.Histological and histomorphometric analysis revealed that the angiogenesis in group D was significantly enhanced as compared with the rest groups(P<0.05).These results strongly suggest that MSCs transfected with rAAV2-bFGF in combination with PDLLA/HA can effectively promote the early angiogenesis of calvarial defects in rats,which played an important role in creating an environment suitable for the survival and activity of transplanted cells for further applications in cranio-maxillofacial bone regeneration.
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To construct the recombinant adeno-associated virus (rAAV) vector with human bone morphogenetic protein 7 (BMP7) and observe the BMP7 mRNA expression in vitro, BMP7 CDS se- quence was cloned into expression plasmid pAAV-MCS of AAV Helper Free System. The recombi- nant plasmid was identified with enzyme digestion and sequencing. The recombinant plasmid, pAAV-RC, pHelper were co-transfected into AAV-293 cells according to the calcium phosphate-based protocol. The viral stock was collected by 4 rounds of freeze/thaw. After purified and concentrated,the recombinant virus titer was determined by dot-blot assay. HEK293 cells were transfected with the recombinant virus at different MOI, and the expression of BMP7 mRNA was detected by RT-PCR. The results showed rAAV-BMP7 was constructed and packaged successfully. The physical particle titer was 2.5×1011 vector genomes/mL. There was different expression level of BMP7 mRNA after transfecton. These data suggested that recombinant AAV mediated a stable expression of hBMP7 mRNA in 293 cells. The AAV production method may pave the way of an effective strategy for the jaw bone defection around dental implants.
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<p><b>OBJECTIVE</b>To introduce a new method for enlarging the survival area of an expanded flap.</p><p><b>METHODS</b>After the skin expander was inflated with enough injection, the first delaying was performed. In the operation, two incisions were made in the skin and subcutaneous tissue superficial to the expander capsule on both sides of the long axis of the expanded flap. After 10 to 14 days, the second delaying followed, in which one pedicle was divided to form a unilateral-pedicled, super-long random flap. When the flap was transferred to the recipient Since 2000,this technique has been used in 16 patients. All the area, the door site was closed directly.</p><p><b>RESULTS</b>flaps survived completely.</p><p><b>CONCLUSIONS</b>The technique of twice delaying can enlarge the survival area of the expanded skin flap.</p>
Subject(s)
Adolescent , Adult , Child , Female , Humans , Male , Cicatrix , General Surgery , Follow-Up Studies , Plastic Surgery Procedures , Methods , Skin Transplantation , Methods , Surgical FlapsABSTRACT
<p><b>OBJECTIVE</b>To investigate hereditary susceptibility to coronary heart disease (CHD) in apolipoprotein E(apo E) and apo B polymorphisms of youths.</p><p><b>METHODS</b>Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to analyze apoE, apoB Xba I, apoB 3' variable number of tandem repeat (VNTR) genotypes for 244 healthy Han students (among them were 109 students with positive CHD family history).</p><p><b>RESULTS</b>The allele frequencies of apo e4, XbaI x(+), 3'VNTR-B(hypervariable element, HVE>38) in the positive group were obviously higher than those in the negative group(P<0.05), and were significantly correlated with the increase in TC, LDL-C, apoB100 levels (P<0.05).</p><p><b>CONCLUSION</b>The alleles for apo e4, XbaI x(+), 3'VNTR-B may be the important genetic markers of Han CHD.</p>