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Objective:To examine the expression of transforming growth factor I(TGF-β1) and bone morphogenetic protein-9 (BMP-9) in human nonunion tissues,and to evaluate the clinical significance.Methods:The number of hypertrophic nonunion tissue samples and atrophic nonunion tissue samples were collected from Department of Orthopedics,the Second Xiangya Hospital of Central South University and Suzhou Kowloon Hospital Affiliated to School of Medicine of Shanghai Jiao Tong University between 2010 and 2014.Semi-quantification of SP immunohistochemical method and pathological image analysis software IPP6.0 were used to analyze the expression of TGF-β1 and BMP-9.Nonunion type,patients' age and nonunion time were statistical analyzed.Results:The absorbance values of TGF-β1 and BMP-9 in the hypertrophic nonunion tissues were 0.3236±0.0390 and 0.1337±0.0400,respectively;while the absorbance values of TGF-β1 and BMP-9 in the atrophic nonunion tissues were 0.3191±0.0369 and 0.1373±0.0423,respectively,with no significant difference between the two types of tissues (both P＞0.05).There was also no significant difference in patients' age and bone nonunion time between them (all P＞0.05).Conclusion:There is no significant difference in osteogenic potential between the hypertrophic nonunion tissues and the atrophic nonunion tissues.
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Objective To evaluate the effectiveness of anticoagulation management by physician-clinical pharmacist team for patients with valvular atrial fibrillation. Methods One hundred and seventy two patients with valvular atrial fibrillation received warfarin therapy for anticoagulation during hospitalization in Linyi People′s Hospital from January 2014 to December 2016, the patients continued to receive warfarin therapy for>6 months after discharge. The patients were randomly assigned in two groups:the anticoagulation management was given by physician-clinical pharmacist team in 87 cases (trial group), while the dosage of wargarin was adjusted in outpatient department by physicians alone in 85 cases (control group). The goal attainment rate of international normalized ratio (INR), the proportion of patients with a stable warfarin dose, knowledge of anticoagulants, belief of medication, medication compliance were compared between two groups. Results There were no significant differences in age, sex, body weight, smoking and drinking habits, valvular disease type, comorbidities; and the initial INR, knowledge of anticoagulants, belief of medication and medication compliance at admission between two groups (all P>0.05). The goal attainment rate of INR (52.17％vs. 41.02％,χ2=8.178, P=0.004), the proportion of patients with a stable dose of warfarin (74.71％ vs. 56.47％,χ2=6.349, P=0.012), the knowledge of anticoagulants (11.03 ± 2.25 vs. 10.08 ± 1.86, t=3.018, P=0.003), the belief of medication[(12.23 ± 2.07) vs. (11.67 ± 1.48), t=2.042, P=0.043], and the medication compliance[(7.36 ± 0.89) vs. (7.04 ± 1.10), t=2.1128, P=0.036] in the trial group were significantly higher than those in control group. Conclusion Anticoagulation management by physician - clinical pharmacist team can improve the management level of anticoagulation and the knowledge of anticoagulans, enhance the medication belief, improve the goal attainment rate of INR and the compliance rate of medication in patients with valvular atrial fibrillation.
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Objective To study the effect of Sarcandra Glabra on the expression of signal transduction molecules of TGF-β1/Smads signaling pathway in miniature pig of radiation-induced lung injury.Methods 75 miniature pigs were divided into control group,radiation group and radiation plus medication group randomly.At 1 week before exposure of right lung with 15 Gy γ-rays,the miniature pigs in radiation plus medication group were given Sarcandra glabra,while those in the other groups received an equal amount of saline.Right lung were taken at weeks 2,4,8,12 and 24 after irradiation,the pathological changes in the lung tissue were observed by HE staining,and the expression of mRNA and protein of TGF-β1,Smad2,Smad3,and Smad7 were detected by real-time PCR and western blotting,respectively.Results Sarcandra glabra reduced the inflammation and fibrosis of the lung tissue in miniature pig after irradiation.Compared with control group,the expressions of TGF-β1 and Smad3 were significantly increased at 2 weeks after irradiation(P ＜ 0.05),Smad2 and Smad7 were increased at 8 and 12 weeks after irradiation(P ＜ 0.05),respectively,in the radiation group.Compared with the radiation group,the expressions of TGF-β1 and Smad2 were significantly decreased(P ＜ 0.05) from the fourth and eighth week,respectively,Smad3 had no obvious change while Smad7 was significantly increased from the second week in the radiation plus medication group (P ＜ 0.05).Conclusions Sarcandra Glabra plays protective effect on radiation-induced lung injury in miniature pig by regulating TGF-β1,Smad2 and Smad7 expressions in the TGF-β1/Smads signaling pathway.
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Objective To explore the influence and mechanism of cytokines and protein expression of alveolar epithelial type (ACE) Ⅱ cells in Bama minipigs' right-thorax with a single 15 Gy dose irradiation.Methods All minipigs received either right thoracic irradiation or sham-irradiation under anesthesia.At 4,8,12 and 24 week post-irradiation,5 minipigs respective and random from irradiarion groups and control group were sacrificed to remove the lungs.The protein expression of surfactant associated protein (SP)-A,transforming growth factor (TGF)-β1,Vimentin and E-cadherin were detected by Western blot.The protein expression of α-smooth muscle actin (SMA) was detected by immunohistochemistry.The co-localization of SP-A and α-SMA was visualized by double immunofluorescence staining.Results At 4,8,12 and 24 week post-irradiation,a significant increase in the protein expression of α-SMA,TGF-β1 and Vimentin were observed in irradiated lung compared to sham-irradiated controls(α-SMA:t =2.46-3.26,P ＜0.05;TGF-β1:t =2.96-3.52,P ＜0.05;Vimentin:t =3.24-5.05,P ＜ 0.05).By contrast,the protein expression of SP-A and E-cadherin in irradiation group was lower than it in control group (SP-A:t =3.62-4.65,P ＜ 0.05;E-cadherin:t =2.53-4.15,P ＜ 0.05).Moreover,at 8 week after irradiation,under confocal laser scanning microscope,the co-localization of SP-A and α-SMA was observed in irradiated alveolar epithelium cells,and it was not observed in sham-irradiated controls.Conclusions These data demonstrate that E-cadherin,SP-A and TGF-β1 may act as sensitive predictors of radiation-induced lung injury(RILI).Irradiation may lead to ACE Ⅱ cells achieving a mesenchymal phenotype,namely,epithelial to mesenchymal cells transition occurs,and ACE Ⅱ cells play the important part in the development of RILI by epithelial-mesenchymal transition.
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BACKGROUND: Diabetic cardiomyopathy, a serious complication of diabetes, is an important factor of increased mortality in patients with diabetes. Therefore, providing an effective experimental animal model is particularly important for studying the pathogenesis and treatment methods of diabetic cardiomyopathy. OBJECTIVE:To explore the method of establishing Wistar rat models of diabetic cardiomyopathy. METHODS:Forty rats were randomly divided into the control group (n=10) and diabetic cardiomyopathy group (n=30). The rats in the diabetic cardiomyopathy group were intraperitonealy injected with 60 mg/kg streptozotocin at a time to establish rat models of diabetic cardiomyopathy. The rats in the control group were given the same dosage of citric acid buffer by the same way. The rats in these two groups were al fed with non-fat high-sugar normal diet. RESULTS AND CONCLUSION:Compared with the control group, after 3 weeks of injection with streptozotocinin in rat models of diabetic cardiomyopathy, blood glucose level was significantly increased, myocardial cels arranged in disorder, the nuclei were of different sizes, colagen content in the myocardial tissue was significantly increased, and colagen fibers were thick and disordered. In addition, the mRNA and protein levels of transforming growth factorβ1 and type I colagen, two indices reflecting myocardial fibrosis, were markedly increased. These results indicate that intraperitonealy injecting large doses of streptozotocin (60 mg/kg) at a time and feeding with non-fat high-sugar normal diet could establish a stable rat model of type 1 diabetic cardiomyopathy. This method is safe and effective with high feasibility.
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Objective To explore the clinical value of serum procalcitonin (PCT)-based antibiotic therapy in the second-classedexacerbations of chronic obstructive pulmonary disease (AECOPD). Methods 240 patients diagnosised as AECOPD were randomized to the PCT group and the control group. Serum PCT levels of patients from the PCT group were measured 1 h after hospitalized and the third, fifth, eighth day respectively. When PCT < 0.1 μg / L, patients will stop taking antibiotics and initiated while PCT≥0.1 μg / L. Antibiotic treatment in the control group was based on guidelines of COPD diagnosis and treatment. Results Duration of antibiotic therapy and hospitalization were respectively 5.6 ± 1.4 and 8.2 ± 1.1 days in the PCT group, 9.2 ± 2.2 and 11.4 ± 2.5 days in the control group (both P < 0.05). Mean costs of hospitalization expensesand antibiotic therapy were 5700 ± 201 and 1650 ± 189) yuan in the PCT group, 6210 ± 220 and 2350 ± 210 yuan in the control group (both P < 0.05). The clinical effective rate, times of exacerbation, one-year ΔFEV1, the 1-year hospitalization rate and time to next exacerbation all showed no significant differences between the two groups. Conclusion PCT-guided antibiotic treatment reduces antibiotic use inthe second-classed acute exacerbations patients.
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Objective To explore the effect of the single maximum of alcohol consumption on vomiting induced by chemotherapy in male patients with nasopharyngeal carcinoma.Methods According to the single maximum of alcohol consumption,48 cases of male patients with nasopharyngeal carcinoma were divided into two groups:group A (small capacity for liquor group 10-50 g),group B (large capacity for liquor group ＞50 g).The amount of alcohol intake was assessed by a questionnaire.The responses of vomiting and the effects of antiemetic therapy between the two groups during the first cycle after induction chemotherapy were observed.Results The incidence rates of vomiting induced by chemotherapy between the two groups were 52.0％ and 17.4％,respectively.There were significant differences between the two groups (x2 =6.273,P ＜0.05).The incidence rates of grade 0,1,2 and 3 of vomiting in group A were 48.0％,20.0％,20.0％,12.0％,in group B were 82.6％,13.0％,4.4％,O,respectively.There were significant differences between the two groups (x2 =6.013,P =0.024).In the two groups,the complete control rate for acute vomit were 48.0％ and 82.6％ (x2 =6.273,P =0.012),for delayed vomit were 36.0％ and 65.2％ (x2 =4.090,P =0.043).There were also significant differences between the two groups.Conclusion With the increasing of capacity for liquor,the incidence of vomiting is significantly reduced,the degree of vomiting is distinctly alleviated,and the antiemetic efficacy is distinctly increased.
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The invasion and metastasis of esophageal carcinoma is associated with several tumor marker on oncogenesi and tumor development,including matrix metalloproteinase,cell adhesion molecules CD44,pituitary-tumour transforming genes.Studies have shown that these tumor markera are over-expression in esophageal carcinoma,which affect the progress and prognosis of esophageal carcinoma,and their specificity and value for clinical application need to be futher studied.
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Local recurrence and distant metastasis is the major factor of impacting long-term survival for patients with esophageal carcinoma after radical resection. Postoperative radiotherapy and chemotherapy is expected to reduce the rate of local recurrence and distant metastasis, but views for the value of postoperative radiochemotherapy were inconsistent in the past. In recent years, scholars have performed much more clinical studies on postoperative radiotherapy and chemotherapy and found that the combined postoperative radiotherapy and chemotherapy after surgery significantly reduces the recurrence rate and distant metastasis rate and can enhance the survival of the patients.
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In order to construct recombinant baculovirus carrying Schistosoma japonicum 26 ku glutathione S-transferase gene (Sj26), and observe the expression of Sj26 in mammalian cells, the Sj26 gene was amplified with plasmid pGEX-3X as template by PCR, and then recombined into T vector for sequencing. Sj26 gene was inserted into the downstream of CMV promoter of donor plasmid pFBDGC, and the recombinant donor plasmid pFBDGC-Sj26 transformed into DH10Bac, then the recombinant bacmid AcCMVSj26 was isolated and transfected into Sf9 cells. The recombinant baculovirus was harvested and final titer of vAcCMVSj26 was measured. BHK cells were transducted with recombinant baculovirus in vitro. By using Western blot, the expression of 26 ku glutathione S-transferase (GST) was detected. The results showed that after enzyme digestion and sequencing, the donor plasmid was successfully constructed. PCR confirmed that pFBDGC-Sj26 and Bacmid homologous recombination occurred in E. coli. After transfection of Sf9 cells with recombinant Bacmid, recombinant baculovirus was replicated in Sf9 cells and expressed green fluorescent protein. PCR further revealed recombinant baculovirus contained Sj26. The titer of the harvested baculovirus was 1.24 x 10(8). Western blot demonstrated that recombinant baculovirus could express 26 ku GST in BHK cells. It was concluded that Sj26 recombinant baculovirus was successfully constructed, and the 26 ku GST was expressed in mammalian cells.
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Objective To compare a potential role of dendritic cells (DCs) and macrophages in inducing protective immunity against infection with Schistosoma japonicum. Methods DCs and macrophages were pulsed in vitro with soluble egg antigen (SEA) of S. japonicum. BALB/c mice were injected three times with DCs or macrophages, either antigen-pulsed or not,and challenged with 40 ± 2 cercariae of S. japonicum per mouse. Worms were collected 42 days later by portal perfusion of the mice and egg number of liver was calculated. To evaluate whether protective immunity had been induced by preparations of DCs or macrophages, the worm burden and fertility ( eggs per female per mouse liver) were compared between the groups of mice. The antibody level against SEA was detected by ELISA. Results With respect to mice injected with untreated cells, numbers of worms and eggs per female worms were significantly reduced in the groups of mice having received pulsed DCs (26. 3% and 37.9%, respectively), or pulsed macrophages (22. 0% and 30.7%). Untreated DCs and macrophages induced no significant effects. The antibody level against SEA rose in sera of all groups of mice up to 42 days after the challenge, but most pronounced in those immunized with pulsed DCs, although this was not significantly different from other groups. Conclusion The results suggest that the protective immunity against S. japonicum might be induced by DCs to a higher extent than by macrophages after in vitro pulsing with egg antigen.
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In order to construct recombinant baculovirus carrying Schistosoma japonicum 26 ku glutathione S-transferase gene (Sj26), and observe the expression of Sj26 in mammalian cells, the Sj26 gene was amplified with plasmid pGEX-3X as template by PCR, and then recombined into Tvector for sequencing. Sj26 gene was inserted into the downstream of CMV promoter of donor plasmid pFBDGC, and the recombinant donor plasmid pFBDGC-Sj26 transformed into DH10Bac,then the recombinant bacmid AcCMVSj26 was isolated and transfected into Sf9 cells. The recombinant baculovirus was harvested and final titer of vAcCMVSj26 was measured. BHK cells were transducted with recombinant baculovirus in vitro. By using Western blot, the expression of 26 ku glutathione S-transferase (GST) was detected. The results showed that after enzyme digestion and sequencing, the donor plasmid was successfully constructed. PCR confirmed that pFBDGC-Sj26 and Bacmid homologous recombination occurred in E. coli. After transfection of Sf9 cells with recombinant Bacmid, recombinant baculovirus was replicated in Sf9 cells and expressed green fluorescent protein. PCR further revealed recombinant baculovirus contained Sj26. The titer of the harvested baculovirus was 1.24 × 108. Western blot demonstrated that recombinant baculovirus could express 26 ku GST in BHK cells. It was concluded that Sj26 recombinant baculovirus was successfully constructed, and the 26 ku GST was expressed in mammalian cells.
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In contemporary studies of pulse and of the relationship between sphygmo-diagram and syndrome types or diseases, pulse instrument is usually applied to trace the sphygmo-diagram. A comparatively systemic theory about pulse diagnosis has been formed, and it promotes the pulse researching process. But the mechanism of pulse is complicated and the expressive information of pulse is diversity. So it is difficult to record the complicated information of pulse by applying the instrument. In addition, the simplicity in methods of tracing and analyzing sphygmo-diagram and the lack of criterion for syndrome differentiation make it difficult to study the relationship between the sphygmo-diagram and the syndrome types. It's important to lay stress on clinical applying, to promote communication among researchers, to unify the standards of pulse instrument and syndrome differentiation, and to reinforce the research on the relationship between sphygmo-diagram and the syndrome types. The government's support is also needed to promote multi-science cooperation at the same time.
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To determine the content of astragaloside IV in Xiao' er Weijining Mixt ure, TCL conditions were optimized by comparing various refined methods. And the y were as follows: the underlayer of CHCl3-CH3OH-H2O(65∶35∶10) below 10 ℃was developed at 15～18℃ and sprayed with 10%H2SO4-C2H5OH,the heating tempe rature was 105℃ for 3～5 min,and λS=400 nm, λR =700 nm.
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Aim Five types of civil Toxoplasma gondii ELISA test kit were used to the detected specimen which proved to be the same, foreign-made test kit was used to examine the positive specimen given by these five types of test kit, as to make comparison about quality of test kit. Results IgG-ELISA A,B kits positive showing rates (sensitive)are all 60％ ,the corresponding rate between A&B is merely 50％ ,the specifics are 35％&90％. C, D,E kits' positive showing rates (sensitive)are 30 ％, 60 ％, 70 ％ the specifice are 75 ％, 54 ％, 87 ％. The result about examining the probes given by A, B, D, E kit :IgG corresponding rates are better, which are all more than 90％ ;IgM corresponding rates are all more than 60％. It turn out that, according to the civil conditions of ELISA test kit, we suggest that besides the quality being improved in, all kinds of test kit should be used combinatively, so as to avoid neglect and mistake on examination, and we should be cautious in examining.
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Objective To study the presence of iNOS in Schistosoma japonicum and demonstrate its distribution in different stages of this schistosome. Methods Cryostat sections for adult worms and paraffin sections for eggs in the liver of infected mouse, sporocysts and cercariae in snails were prepared, immunofluorescent test was performed to detect the presence of iNOS in adult worms, sporocysts, cercariae and miracidium, the distribution of this enzyme was observed in different stages of Schistosoma japonicum. Western blotting was used to further demonstrate the immunoreactivity to iNOS in adult worms. Results The results of immunofluorescent test showed that specific yellow- green fluorescence was mainly among subtugment of adult worms. Positive staining was also distributed on the surface of miracidium and its glands. For both sporocysts and cercariae, the majority of fluorescence was on their surface. Anti-iNOS antibody could recognize an apparent specific band in Western blotting of adult worm proteins, with a of Mr 210 000. Conclusion There is an expression of iNOS-like enzyme in Schistosoma japonicum.
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Objective To explore possible associations between host polymorphism of HLA classⅡgenotypes and advanced hepatosplenic schistosomiasis japonica.Methods 45 advanced schistosomiasis patients(experimental group) and 44 age,and sex,matched patients with chronic schistosomiasis(control group) from the same area were investigated for their HLA class II gene DRB genotypes by genotyping the alleles using microarray DNA chip.The correlation of allele frequencies to advanced hepatosplenic schistosomiasis was compared for the two groups.Results HLA,DRB1*04x exhibited markedly higher frequency in advanced patients than that in control group(P
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Objective To compare the immune protective effect of different antigens of Trichinella spiralis:excretory-secretory(ES) antigen of adult worms, ES antigen of muscle larva and their mixed antigens in mice. Methods ES antigens of Trichinella spiralis adult worms and muscle larva were produced by the physiological saline culture methods. The ES antigens of Trichinella spiralis adult worms, muscle larva and the mix were used to immunize mice 3 times at 7 day interval, adjuvant control and normal control were set up. Seven day after the final immunization, each mouse was orally challenged with 200 Trichinella spiralis larva. Intestinal adult worms and muscle larva of Trichinella spiralis of each group were recoveried and examined on Day 7 and Day 30 post-challenge,respectively. Results The intestinal adult worms of Trichinella spiralis reduced by 87.95%, 69.48% and 84.34% in the adult worm ES antigen group, muscle larval ES antigen group and their mixed antigen group, respectively, while muscle larva reduced by 74.79%, 87.97% and 86.87% respectively. Adult worm reduction rates of the adult worm ES antigen group and mixed antigen group were higher than those of muscle larval ES antigen group (P
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Objective To investigate the effect of nitric oxide on the liver fibrosis of mice infected with Schistosoma japonicum during the early stage of schistosomiasis. Methods NMRI mice were treated with AMG-an iNOS inhibitor from day 23 post infection(p.i) to the sacrificed and the livers were collected at 38 days, 45 days p.i respectively. The expression and distribution of collagen typeⅠ, Ⅲ (ColⅠand ColⅢ) in liver tissues were investigated with the Picric acid-Sirius red staining techniques and differentiated with the polarization microscopy combining with picture analysis system. Hydroxyproline concentration in liver homogenate was measured by the biochemical methods. Results At 38 day p.i, the Picric acid-Sirius red staining showed that the hyperplasia of Col Ⅰ and ColⅢ in the livers of AMG treated mice increased significantly compared with the control livers, but there was no significant difference of hydroxyproline content between the two groups. At 45 days p.i, only the hyperplasia of Col Ⅰ in AMG treated group increased significantly compared with the control livers, and moreover, content of hydroxyproline of the inhibited mice was significantly higher than that of the control mice (P
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Objective To explore the mechanism of protective immunity against Schistosoma japonicum infection induced by Sj26 gene transfected dendritic cell(DC).Methods 48 BALB/c mice were divided randomly into 4 groups with 12 each.The mice were injected through auricle for three times with Sj26 gene transfected DC(Group A),pcDNA3 transfected DC(Group B),untreated DC(Group C) and RPMI-1640(Group D) respectively,and challenged with 40?2 cercariae of S.japonicum per mouse 2 weeks after the last immunization.Sera from mice were examined for IgG antibody,IFN-? and IL-4 by ELISA.Western blot was used for detecting specific anti-Sj26 IgG antibody.The production of IFN-? and IL-4 in the supernatant of spleen cells stimulated with soluble egg antigen(SEA) and ConA was quantified by sandwich ABC-ELISA.The proliferation of spleen cells were measured with MTT method.Results IgG antibody increased significantly in the mice of group A at 2 weeks after the last immunization(absorbency A491=0.117),higher than that of group B(A491=0.061) and group C(A491=0.058)(P