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Itch is an unpleasant sensation that provokes the desire to scratch. While acute itch serves as a protective system to warn the body of external irritating agents, chronic itch is a debilitating but poorly-treated clinical disease leading to repetitive scratching and skin lesions. However, the neural mechanisms underlying the pathophysiology of chronic itch remain mysterious. Here, we identified a cell type-dependent role of the anterior cingulate cortex (ACC) in controlling chronic itch-related excessive scratching behaviors in mice. Moreover, we delineated a neural circuit originating from excitatory neurons of the ACC to the ventral tegmental area (VTA) that was critically involved in chronic itch. Furthermore, we demonstrate that the ACC→VTA circuit also selectively modulated histaminergic acute itch. Finally, the ACC neurons were shown to predominantly innervate the non-dopaminergic neurons of the VTA. Taken together, our findings uncover a cortex-midbrain circuit for chronic itch-evoked scratching behaviors and shed novel insights on therapeutic intervention.
Subject(s)
Mice , Animals , Gyrus Cinguli/physiology , Pruritus/pathology , Mesencephalon , Cerebral Cortex/pathology , Neurons/pathologyABSTRACT
Objective: To observe the clinical efficacy modified Shenling Baizhu San in treating patients with spleen deficiency wetness type subacute eczema, and its effect on the inflammatory factors and peripheral blood T lymphocyte subsets, so as to provide a theoretical basis for clinical treatment. Method: Totally 176 cases of spleen deficiency wetness type subacute eczema treated at our hospital from Jan. 2016 to Dec. 2017 were selected and randomly divided into control group (88 cases) and observation group (88 cases). Control group was treated with levocetirizine hydrochloride tablets and compound econazole nitrate gel, and observation group was given modified Shenling Baizhu San in addition to therapy of control group for 4 weeks. The clinical efficacy, pruritus score, severity index(EASI)score, dermatology quality of life scales(DQOLS) score and recurrence rate were observed,and inflammatory factor and peripheral blood T lymphocyte subgroup level were detected. Result: After treatment, the pruritus score and EASI score in observation group were lower than those before treatment (PPPPβ, IL-2, IL-4, IL-5 and tumor necrosis factor(TNF) -α in observation group were lower than those before treatment (P+ and CD4+/CD8+ were higher than those of before treatment, and significantly higher than those of control group (PPConclusion: Modified Shenling Baizhu San could enhance the clinical efficacy by inhibiting the inflammatory reaction and regulating the immune function of patients.
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Objective:To observe the effect of Wendantang on tumor necrosis factor-α (TNF-α),interleukin(IL)-6,IL-17,IL-22 and other related inflammatory factors in peripheral blood and the expression of STAT3[the key molecule of Janus kinase 2/signal transducer and activator of transcription 3(JAK2/STAT3) signal pathway in hypothalamus] mRNA and protein of obese rats with syndrome of phlegm-dampness,so as to explore the internal mechanism of Wendantang in interfering obesity with syndrome of phlegm-dampness. Method:A total of 100 rats were randomly divided into blank group(30 rats) and modeling group(70 rats),rats in the blank group was fed with basic feed and the modeling group was fed with high-fat feed for 6 weeks.Animal model of obesity with syndrome of phlegm-dampness was established by the method in literature.After successful modeling,16 obese rats were selected and randomly divided into the model group and Wendantang intervention group with 8 rats in each group,and another 8 rats in the blank group were randomly selected as the normal group.Rats in Wendantang intervention group were given 15 g·kg-1 of crude drug by gavage,while the model group and the normal group were given the same amount of distilled water for gavage once a day for 6 weeks.No eating but no prohibiting drinking before dealing with 12 h and then taking samples after anesthesia.The body weight,Lee's index and obesity rate of rats were measured,the levels of blood lipids[total cholesterol(TC),triglyceride(TG),low density lipoprotein cholesterol(LDL-C) and high density lipoprotein cholesterol(HDL-C)] of rats were detected with a full-automatic biochemical analyzer according to the requirements of the kit,the expression of TNF-α,IL-17,IL-22 and IL-6 in peripheral blood of rats was detected with enzyme-linked immunosorbent assay(ELISA),STAT3 mRNA expression in hypothalamus of rats was detected with real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) and the expression of STAT3 protein in hypothalamus of rats was detected with Western blot. Result:The high-fat feed feeding could successfully replicate the obese rat model,and the obesity rate of rats in the modeling group was greater than 20%,and compared with the blank group,the body weight and Lee's index of rats in the modeling group were significantly increased(PPPPPPPPPPPConclusion:Wendantang has a good effect on improving phlegm-dampness in obese rats,and the mechanism may be related to regulating JAK2/STAT3 signal pathway then to improve the chronic inflammatory state of the body,and all of which provides a scientific basis for Wendantang in intervening obesity with syndrome of phlegm-dampness.
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Ontology construction is a new method for knowledge organization in the background of semantics. The basic method and procedure for ontology construction for channel and acupuncture point therapy were described, the role of ontology in semantic relationship and added knowledge organization value was assayed according to the retrieval, rationalization and visualization demonstration by making use of the ontology construction software, which showed that the method for ontology construction is characterized by its wide application range, deep application level, semantic enrichment and strong relevance. It is convenient for knowledge reorganization and can thus provide a new angle for scientific research and create a new value.
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<p><b>Objective</b>To study the expression of CLAUDIN-11 in the testis tissue of non-obstructive azoospermia (NOA) patients with different severities and investigate its clinical significance.</p><p><b>METHODS</b>Sixty-two NOA patients were divided into a hypospermatogenesis (HS) group (n = 30) and a Sertoli cell only syndrome (SCO) group (n =32). The expression of CLAUDIN-11 in the testicular tissue of the patients was detected by immunohistochemistry, that of CLAUDIN-11 mRNA determined by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR), and the levels of serum reproductive hormones measured by chemiluminescent immunoassay.</p><p><b>RESULTS</b>Immunohistochemistry showed that the expression of CLAUDIN-11 was mainly in the cytoplasm of the Sertoli cells around the seminiferous tubule wall in the HS group, but diffusely distributed in the membrane of the Sertoli cells in the SCO group. RT-qPCR revealed a significantly lower expression of CLAUDIN-11 mRNA in the HS than in the SCO group (0.008 ± 0.001 vs 0.013 ± 0.002, t = 10.616, P<0.01). The level of serum luteotropic hormone (LH) was also markedly lower in the HS than in the SCO group (［3.62 ± 1.34］ vs ［4.96 ± 3.10］ IU/L, P<0.05) and so was that of follicle-stimulating hormone (FSH) (［5.36 ± 2.80］ vs ［10.65 ± 9.18］ IU/L, P<0.05).</p><p><b>CONCLUSIONS</b>The up-regulated expression of CLAUDIN-11 in Sertoli cells may play an important role in the development and progression of spermatogenic dysfunction in NOA patients.</p>
Subject(s)
Humans , Male , Azoospermia , Genetics , Metabolism , Claudins , Metabolism , Follicle Stimulating Hormone , Metabolism , Oligospermia , Genetics , Metabolism , RNA, Messenger , Metabolism , Seminiferous Tubules , Metabolism , Sertoli Cell-Only Syndrome , Genetics , Metabolism , Sertoli Cells , Metabolism , Spermatogenesis , Testis , MetabolismABSTRACT
AIM:To evaluate the therapeutic effects of the fine sight training with the smartphones and pads on hyperopia amblyopia of children.METHODS:One hundred and twenty children (120 eyes) with hyperopia amblyopia were randomly divided into two groups in this prospective study.All the children in these two groups received the basic treatments of spectacle correction,penalization therapy and amblyopia trainings.The treatments of red-light blinking and grating as well as traditional fine sight training were used for the children in the control group.However,the smartphones and pads were applied instead of the traditional performances for the fine sight training in the experimental group.Best corrected visual acuity of every child was tested for every 3mo,to observe the time for the visual improvement and efficacy.RESULTS:In comparison with the control group,significant shorter time (80.54 ± 30.87d,P＜ 0.05) for promoted one line of LogMAR visual acuity and average treatment time (15.34±7.24mo,P＜0.05) were harvested in the experimental group.The efficacy in the experimental group was significantly higher than that in the control group (Z=-2.37,P=0.02).CONCLUSION:The fine sight training with the smartphones and pads can improve vison faster than traditional methods and decrease the time of therapy in children with hyperopia amblyopia,thus providing a new strategy for the treatment of hyperopia amblyopia.
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To apply preventive T-shaped enterostomy in protective defunctioning stoma.Technique of enterostomy closure was only made in abdominal wall rather than abdominal cavity.A total of 78 patients undergoing protective ostomy from January 2005 to April 2012 were divided into two groups of routine enterostomy (group A,n =52) and protective T-shaped enterostomy (group B,n =26).The length of operation and average hospital stay in group A with stoma closure were(98 ± 16) min and (15.3 ± 5.2)days while those of group B (52 ± 9) min and (9.5 ± 2.3) days.The inter-group differences were statistically significant respectively (P ＜ 0.05).Therefore protective T-shaped enterostomy,showing advantages in operative skills,operative trauma and postoperative recovery time,is an effective technique and its application should be further promoted.
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<p><b>OBJECTIVE</b>To discuss the longterm quality-of-life related to swallowing function after different partial laryngectomy.</p><p><b>METHODS</b>The worldwide known swallow quality-of-life questionnaire (SWAL-QOL, Hongkong, Chinese edition); was used in this research to evaluate the swallowing QOL on 96 postoperative patients who underwent different kinds of partial laryngectomy more than one year before. The patients were divided into 4 groups: supracricoid partial laryngectomy-cricohyoidopexy (SCPL-CHP), supracricoid partial laryngectomy-cricohyoidoepiglottopexy (SCPL-CHEP), horizontal supraglottic partial laryngectomy (horizontal PL), and vertical partial laryngectomy (vertical PL).</p><p><b>RESULTS</b>A one-way MANOVA revealed a significant multivariate (the 11 scales of SWAL-QOL) main effect for groups (P < 0.01), and significant univariate main effects were obtained for groups in 9 scales out of 11 (P < 0.01). In all the 9 scales vertical PL group acquired near full scores except the communication scale, and was significantly higher than the other 3 groups in many scales (P < 0.05). CHP group acquired the worst scores of the 4 groups, showing significant differences in most of the 9 scales (P < 0.05). No significant difference was found between Horizontal PL and CHEP except in communication (P > 0.05). Patients with deglutition disorders (choke/cough) had a lower score in the social function scale.</p><p><b>CONCLUSIONS</b>Swallowing quality-of-life of postoperative patients was deeply influenced even when more than one year had passed after surgery. Some of them felt deeply burdened by deglutition disorder. Patients after CHP proved to have a worst quality of life than the others, while vertical PL the best. The QOL between Horizontal PL and CHEP was shown to be almost the same. The influence over QOL from longterm dysphagia was multi-dimensional, containing the degeneration of social function.</p>
Subject(s)
Aged , Aged, 80 and over , Humans , Male , Middle Aged , Carcinoma, Squamous Cell , General Surgery , Deglutition , Deglutition Disorders , Epidemiology , Laryngeal Neoplasms , General Surgery , Laryngectomy , Methods , Quality of Life , Surveys and Questionnaires , Treatment OutcomeABSTRACT
<p><b>OBJECTIVE</b>To Evaluate the incidence rates and extents of deglutition disorder in patients with laryngeal carcinoma after different types of supracricoid partial laryngectomy.</p><p><b>METHODS</b>Retrospective analysis of postoperative deglutition disorder in patients with laryngeal carcinoma after different types of supracricoid partial laryngectomy treated in our department from 2005 to 2009. The extents of postoperative deglutition disorder were evaluated using a homemade quantitative score table at 5-20 days postoperation.</p><p><b>RESULTS</b>The score of deglutition disorder was 2.71 ± 0.31 in the supracricoid partial laryngectomy-cricohyoidoepiglottopexy (SCPL-CHEP) operation group and 3.43 ± 0.64 in the supracricoid partial laryngectomy-cricohyoidopexy (SCPL-CHP) group, respectively. The deference was statistically significant between the two groups (P < 0.001). The coefficient between age and score of postoperative deglutition disorder was assessed by Pearson correlation analysis. The coefficient of correlation was 0.947 (P < 0.0001) in the SCPL-CHEP group and 0.907 (P < 0.0001) in the SCPL-CHP group. The incidence rate of deglutition disorder was 1/37 in the SCPL-CHEP group and 7/30 in the SCPL-CHP group, evaluated at 8 weeks postoperation (P = 0.012). The deference between the two groups was significant.</p><p><b>CONCLUSIONS</b>The type of operation procedure is an important factor affecting the occurrence of postoperative deglutition disorder in the patients after supracricoid partial laryngectomy, more serious in the SCPL-CHP group. The severity of postoperative deglutition disorder is more serious along with the increase of patient's age. For the elderly (> 70 years of age) patients with laryngeal carcinoma, the choice of surgical procedure should be more cautious, especially with the SCPL-CHP operation.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Age Factors , Cricoid Cartilage , General Surgery , Deglutition Disorders , Laryngeal Neoplasms , General Surgery , Laryngectomy , Methods , Postoperative Period , Retrospective StudiesABSTRACT
Clinical data of 14 patients with iatrogenic duodenal injuries treated in hospital from January 2000 and January 2010 were retrospectively reviewed.Iatrogenic duodenal injuries were found intraoperatively in 9 cases,in whom repair or additional jejunostomy was performed and all were cured and discharged.In 2 patients the duodenal injuries were found within 24 hours postoperatively,1 was cured,another had low flow duodenal fistula and cured with conservative treatment.Duodenal bypass and extraoral drainage were performed in 2 patients whose duodenal injuries was found 72 hours after surgery and died from severe infection of retroperitoneal space and multiple organ failure respectively.One patient whose duodenal injury was found 7 d postoperatively suffered from septic shock and died in 4 h after admission.The results suggest that early detection and early management would result in satisfied outcome for patients with iatrogenic duodenal injuries,the first 24 hours are crucial.
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ObjectiveTo observe the effect of succus entericus reinfusion with continuous enteral nutrition on the barrier function of intestinal mucosa and nutritional status in patients with stomal type fistulas. Methods Sixteen patients with stomal type fistula from July 1995 to May 2008 were enrolled in the study. A]l patients met the following conditions: gut function returned normal; abdominal infection was controlled; total enteral nutrition was provided ; and the length of small intestine for succus entericus reinfusion was more than 50 cm. Intestinal mucosa was taken at 25 to 30 cm away from stoma of fistula by endoscope 0, 7, and 14 days after reinfusior. Hematoxylineosin staining was performed to count the number of intestinal intraepithelial lymphocytes (IIELS). In addition,proliferating cell nuclear antigen (PCNA) was measured with immunohistochemical staining. Serum protein levels were determined by immunonephelometry. ResultsThe percentage of IIELS in intestinal mucosa ( 19.06％ ±4.81％ vs. 12.81％ ±2.95％, P=0.000) and the percentage of PCNA positive cells ( 12.13％ ±4.33％ vs.6.44％ ± 2.34％, P =0.000) 14 days after succus entericus reinfusion were significantly higher than those on the day of reinfusion. Serum fibronectin level increased from ( 152.80 ± 16.50 ) to ( 227.05 ± 45.36 ) mg/L ( P =0.000), and transferring protein level increased from ( 2.16 ± 0.52 ) to ( 2.62 ± 0.41 ) g/L ( P =0.017 ) 14days after succus entericus reinfusion. ConclusionSuccus entericus reinfusion is effective in protecting the intestinal mucosa in patients with stomal type fistulas.
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ObjectiveTo investigate the amount of bacteria and the expression of amylase and lipases in the drainage fluid in patients with intestinal fistulas with time courses.MethodsThe samples were collected from 16 patients with high intestinal fistulas from July 1998 to January 2008.The amounts of bacteria from the drainage fluid were measured 0,2 and 4 hours after taking out from the patients.At the respective time points,the intestinal juices were also collected to measure the amylase and lipase expressions.After reinfusion of succus entericus,thelevels of albumin,prealbumin,transferring,and fibronectin were measured at 0,7,and 14 days,ResultsThere was no significant increase of bacteria in the drainage fluid within 4 hours ( F(0,2) =18 812.50,P ＞ 0.05 ; F(0,4) =387 625.00,P ＞ 0.05).and there was no change in the expressions of amylase ( F(0,2) =190.60,P ＞ 0.05 ;F(0,4) =631.75,P＞0.05) and lipase within 4 hours (F(0,2) =204.10,P＞0.05; F(0,4) =1080.05,P＞0.05).After succus entericus reinfusion,the fibronectin (F(0,14) =74.24,P ＜ 0.01 ; F(7,14) =59.78,P ＜0.01),transferring (F(0,14) =0.46,P ＜ 0.01 ; F(7,14) =0.39,P ＜ 0.05 ),and prealbumin ( F(0,14) =54.37,P ＜ 0.05) were increased significantly.ConclusionsBacteria and enzymes do not increase in the drainage fluid within 4 hours in patients with intestinal fistulas.Therefore,it is safe and effective to reinfuse succus entericus.
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Objective To evaluate the biomechanical effects of using a new injectable calcium phosphate cement to consolidate the fixation of osteoporotic intertrochanteric fracture.Method Five matchod pairs of human cadaver femora were used to produce the model of intertrochanteric fracture.All fractures were fixed with dynamic hip screws(DHS),and divided into two groups.In the CPC consolidation group of each pair,CPC was used to grout the hip screw and to fill the posteromedial defect.All femora were subjected to biomechanical test.Results Under the loading of 500 N,in the CPC consolidation group,the mean axial stiffness is(691.93±18.90)N/mm and the horizontal shear stiffness is(5553.84±27.47)N/mm.The mean lateral and medial strength is(5.15±0.35)MPa and(4.13±0.24)MPa.The torsion stiffness was 0.41 and the ultimate loading is(3580±286)N.In the control group,the mean axiak stiffness is(453.45±19.75)N/mm,the horizontal shear stiffness is(3848.87±22.63)N/mm,the mean lateral and medial strength is(3.12±0.37)MPa and(1.80±0.21)MPa,and,the torsion stiffness is 0.35 and the ultimate loading is(2512±189)N.Consolidation fixation with CPC increased each of the biomechanical efficiency(P＜0.05).Conclusions CPC consolidation of osteoprotic femoral head and the medial defect of intertrochanteric fracture can significantly improve the overall stability and decrease the rate of postoperative complication.
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Objective To evaluate the biomechanical effects of using a new injectable calcium phosphate cement to consolidate the fixation of osteoporotic intertrochanteric fracture.Method Five matchod pairs of human cadaver femora were used to produce the model of intertrochanteric fracture.All fractures were fixed with dynamic hip screws(DHS),and divided into two groups.In the CPC consolidation group of each pair,CPC was used to grout the hip screw and to fill the posteromedial defect.All femora were subjected to biomechanical test.Results Under the loading of 500 N,in the CPC consolidation group,the mean axial stiffness is(691.93±18.90)N/mm and the horizontal shear stiffness is(5553.84±27.47)N/mm.The mean lateral and medial strength is(5.15±0.35)MPa and(4.13±0.24)MPa.The torsion stiffness was 0.41 and the ultimate loading is(3580±286)N.In the control group,the mean axiak stiffness is(453.45±19.75)N/mm,the horizontal shear stiffness is(3848.87±22.63)N/mm,the mean lateral and medial strength is(3.12±0.37)MPa and(1.80±0.21)MPa,and,the torsion stiffness is 0.35 and the ultimate loading is(2512±189)N.Consolidation fixation with CPC increased each of the biomechanical efficiency(P＜0.05).Conclusions CPC consolidation of osteoprotic femoral head and the medial defect of intertrochanteric fracture can significantly improve the overall stability and decrease the rate of postoperative complication.
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Objective Objective To evaluate the biomechanical effects of using a new injectable calcium phosphate cement to consolidate the fixation of osteoporotic intertrochanteric fracture. Method Five matched pairs of human cadaver femora were used to produce the model of intertrochanteric fracture. All fractures were fixed with dynamic hip screws(DHS),and divided into two groups. In the CPC consolidation group of each pair, CPC was used to grout the hip screw and to fill the posteromedial defect. All femora were subjected to biomechanical test. Result Under the loading of 500 N, in the CPC consolidation group, the mean axial stiffness was (691.93±18.90) N/mm and the horizontal shear stiffness was (5553.84±27.47) N/mm. The mean lateral and medial strength was 5.15±0.35 MPa and (4.13±0.24) MPa. The torsion stiffness was 0.41 and the ultimate loading was (3580±286)N. In the control group, the mean axial stiffness was (453.45±19.75) N/mm, the horizontal shear stiffness was (3848.87±22.63) N/mm, the mean lateral and medial strength was (3.12±0.37) MPa and (1.80±0.21) MPa, and, the torsion stiffness was 0.35 and the ultimate loading was (2512±189)N. Consolidation fixation with CPC increased each of the biomechanical efficiency（P<0.05）. Conclusion CPC consolidation of osteoprotic femoral head and the medial defect of intertrochanteric fracture could significantly improve the overall stability and decrease the rate of postoperative complication.
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<p><b>OBJECTIVE</b>To compare the results of extended vertical partial laryngectomy (similar to modified supracricoid partial laryngectomy with cricohyoidoepiglottopexy) and cricohyoidoepiglottopexy in the treatment of laryngeal carcinoma.</p><p><b>METHODS</b>Retrospectively analyzed on the results and prognosis in patients underwent extended vertical partial laryngectomy and cricohyoidoepiglottopexy between 1998 and 2005. The operation was similar to extended vertical partial laryngectomy. The healthy vocal cord and ventricular band as well as about 1/3 to 2/3 laminas of thyroid cartilage were removed. The healthy cricoarytenoid joint was reserved. The vocal cord, ventricular band, fixed or limitation of motion arytenoid cartilage and 2/3 laminas of thyroid cartilage in ill side were removed. The posteroinferior border of laminas of thyroid cartilage in both sides were reserved. The cricoid was lifted and fixed with hyoid epiglottis directly. Extended vertical partial laryngectomy group consisted of 37 patients with glottic carcinoma (stage T2 16 cases, stage T3 21 cases) and cricohyoidoepiglottopexy group consisted of 34 patients with glottic carcinoma (stage T2 12 cases, stage T3 21 cases, stage T4 1 case).</p><p><b>RESULTS</b>Kaplan-Meier analysis was performed to calculate the survival rates. The three-year cumulative survival rate was 91.7% in extended vertical partial laryngectomy group and 87.5% in cricohyoidoepiglottopexy group respectively. There was no significant difference between the two groups (P > 0.05). The five-year cumulative survival rate was 80.6% in extended vertical partial laryngectomy group and 81.3% in cricohyoidoepiglottopexy group respectively. There was also no significant difference between the two groups (P > 0.05). The decannulation rate was 100% (37/37) in extended vertical partial laryngectomy group and 94.1% (32/34) in cricohyoidoepiglottopexy group respectively. The decannulation time was (14.0 + or - 2.3) days in extended vertical partial laryngectomy group and (19.0 + or - 4.6) days in cricohyoidoepiglottopexy group respectively. The incidence of aspiration was 2.7% (1/37) in modified group and 23.5 (8/34) in cricohyoidoepiglottopexy group respectively evaluated at 8th weeks post-operatively. The evaluation of deglutition disorder was analyzed by Ridit analysis in both groups and the results showed that there was significant difference between the two groups (U = 7.341, P < 0.001). The symptom of aspiration in extended vertical partial laryngectomy group was significant less than in cricohyoidoepiglottopexy group.</p><p><b>CONCLUSIONS</b>Although the survival rate was not different between the two groups. The preservation of laryngeal function in extended vertical partial laryngectomy group was significant better than in cricohyoidoepiglottopexy group and extended vertical partial laryngectomy.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Cricoid Cartilage , General Surgery , Hyoid Bone , General Surgery , Laryngeal Neoplasms , General Surgery , Laryngectomy , Methods , Prognosis , Retrospective Studies , Survival RateABSTRACT
Influenza A/H1N1 virus-encoded nonstructural, or NS1, protein inhibits the 3'-end processing of cellular pre-mRNAs by binding the cellular protein: the 30-kDa subunit of CPSF (cleavage and polyadenylation specificity factor, CPSF30). CPSF30 binding site of the NS1 protein is a potential target for the development of drugs against influenza A/H1N1 virus. A yeast two-hybrid screening system was constructed and used for screening Chinese medicines that inhibit the interaction of the A/H1N1 flu NS1 protein and human CPSF30 protein. The NS1 gene of A/H1N1 virus was amplified by consecutive polymerase chain reaction (PCR), and the human CPSF30 gene of HeLa cell cloned by reverse transcriptase-polymerase chain reaction (RT-PCR). Then the two gene fragments confirmed by sequencing were subcloned into the yeast expression vectors pGBKT7 and pGADT7, respectively. The two constructs, bait vector pGBKNS1 and prey vector pGADCPSF, were co-transformed into yeast AH109. The eight individual yeast colonies were picked and subjected to verification by PCR/gel electrophoresis. The inhibition of the NS1-CPSF30 interaction was allowed the identification of selective inhibitors. The four of more than thirty identified Chinese medicines, including 'Shuanghuanglian oral liquid', showed the strong inhibition of the NS1-CPSF30 interaction.
Subject(s)
Humans , Base Sequence , Binding Sites , Cleavage And Polyadenylation Specificity Factor , Genetics , Metabolism , Drugs, Chinese Herbal , Pharmacology , Gene Amplification , HeLa Cells , Influenza A Virus, H1N1 Subtype , Genetics , Peptide Fragments , Genetics , Plasmids , Protein Binding , Transformation, Genetic , Two-Hybrid System Techniques , Viral Nonstructural Proteins , Genetics , MetabolismABSTRACT
Objective To evaluate the biomechanical effects of using a new injectable calcium phosphate cement to consolidate the fixation of osteoporotic intertrochanteric fracture.Method Five matchod pairs of human cadaver femora were used to produce the model of intertrochanteric fracture.All fractures were fixed with dynamic hip screws(DHS),and divided into two groups.In the CPC consolidation group of each pair,CPC was used to grout the hip screw and to fill the posteromedial defect.All femora were subjected to biomechanical test.Results Under the loading of 500 N,in the CPC consolidation group,the mean axial stiffness is(691.93±18.90)N/mm and the horizontal shear stiffness is(5553.84±27.47)N/mm.The mean lateral and medial strength is(5.15±0.35)MPa and(4.13±0.24)MPa.The torsion stiffness was 0.41 and the ultimate loading is(3580±286)N.In the control group,the mean axiak stiffness is(453.45±19.75)N/mm,the horizontal shear stiffness is(3848.87±22.63)N/mm,the mean lateral and medial strength is(3.12±0.37)MPa and(1.80±0.21)MPa,and,the torsion stiffness is 0.35 and the ultimate loading is(2512±189)N.Consolidation fixation with CPC increased each of the biomechanical efficiency(P＜0.05).Conclusions CPC consolidation of osteoprotic femoral head and the medial defect of intertrochanteric fracture can significantly improve the overall stability and decrease the rate of postoperative complication.
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Objective To develop a Simple,accurate,rapid,economic,large-scale detection method for the detection of singe nucleotide polymorphisms (SNPs) metabolic enzymes,using polymerase chain reaction with confronting two-pair primers (PCR-CTPP).Methods The primers of CYP1A1 (A4889G),EPHX1 (A416G) and NQO1 (C609T) were designed for PCR-CTPP,and the PCR conditions were optimized.The results of genotyping were verified by DNA sequencing.The above SNPs were detected by the PCR-CTPP detection method in a randomly selected 183 healthy individuals of Han ethnicity.The genotype frequencies were analyzed and compared with people from other ethnicities.Results The allele-specific bands of CYP1A1 (A4889G),EPHX1 (A416G) and NQO1 (C609T) were successfully amplified by PCR-CTPP under the optimal conditions and the results of genotyping were consistent with DNA sequencing.Among 183 healthy Han individuals,the genotypic distributions of CYP1A1 (A4889G),EPHX1 (A416G) and NQO1 (C609T) showed that the wild-type,homozygous variants,and heterozygotes were 103 (56.3%),8 (4.4%),72 (39.3%) and 142 (77.6%),4 (2.2%),37(20.2% ),60(32.8% ),32 (17.5%),91 (49.7%) respectively.The distributions of genotypes were all in accordance with the Hardy-Weinberg equilibrium (P＞0.05),with statistical differences and with other ethnic populations(P＜0.05).Conclusion The SNPs of metabolic enzymes can be detected by PCR-CTPP method which is simple,accurate,rapid,economic and with large scale.PCR-CTPP can be used for large scale clinical and epidemiological screening.
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<p><b>OBJECTIVE</b>To investigate the specific anti-breast cancer immune response induced by dendritic cells (DC) loaded with trastuzumab and apoptotic Her-2+ breast cancer cells.</p><p><b>METHODS</b>DCs were generated from healthy peripheral blood mononuclear cells (PBMCs) in the presence of recombinant cytokines GM-CSF, IL-4 and TNF-alpha. Mature DCs were harvested after 7 days' co-culture of PBMCs and trastuzumab-treated apoptotic SKBr3 cells. The morphologic characteristics and ultrastructure of the DC were observed under the inverted phase-contrast microscope and transmission electron microscope (TEM), respectively. Flow cytometry (FCM) was used to check the expression of several DC specific markers: CD14, CD1a, CD64, CD80, CD83, CD86, HLA-ABC and HLA-DR. DC-cytokine induced killer (DC-CIK) cells were prepared by co-culture of DCs and peripheral blood lymphocytes in the presence of anti-CD3 antibodies and human IL-2 at an appropriate concentration. The number of antigen-specific T cells was analyzed by human interferon gamma enzyme linked immunospot (ELISPOT) assay. MTT assay was employed to assess the lysis of breast cancer cell line induced by DC-CIK cells.</p><p><b>RESULTS</b>5 minutes after the adding of DCs to SKBr3 cells pretreated with trastuzumab, the apoptotic SKBr3 cells were found to be circled by DCs. 48 hours later, many membrane-wrapped organelles of the apoptotic target cells in the cytoplasm of DCs were found by TEM. The majority of the organelles were degraded. Fewer organelles from the apoptotic cells were found in DCs without Herceptin. More than 60% in every group of DCs expressed a high-affinity receptor for IgG (FcgammaRI or CD64). CD14 expression on the mature DCs were comparatively lower, and HLA-DR and HLA-ABC expressions were higher in the trastuzumab group. The expression of CD1a, CD80, CD83 and CD86 in trastuzumab group were higher than those in immature DCs group (P < 0.05). ELISPOT assay suggests that the spot number of antigen-specific T cells were higher in trastuzumab group than that in the antigen unloaded DCs group (P < 0.05). The lysis of SKBr3 cells induced by the SKBr3 antigen loaded DC-CIK cells were 1.7 times higher than that of CIK.</p><p><b>CONCLUSION</b>The lysis of SKBr3 cells induced by DC-CIK was increased after that DCs were combined with trastuzumab to capture antigen from SKBr3 cells. These findings support further investigation into the use of combination immunotherapy of the humanized monoclonal antibody, DC vaccines and immunological effector cells.</p>