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OBJECTIVES@#This study aims to investigate the effects of tumor-stromal fibroblasts (TSFs) on the proliferation, invasion, and migration of salivary gland pleomorphic adenoma (SPA) cells in vitro.@*METHODS@#Salivary gland pleomorphic adenoma cells (SPACs), TSFs, and peri-tumorous normal fibroblasts (NFs) were obtained by tissue primary culture and identified by immunocytochemical staining. The conditioned medium was obtained from TSF and NF in logarithmic phase. SPACs were cultured by conditioned medium and treated by TSF (group TSF-SPAC) and NF (group NF-SPAC). SPACs were used as the control group. The proliferation, invasion, and migration of the three groups of cells were detected by MTT, transwell, and scratch assays, respectively. The expression of vascular endothelial growth factor (VEGF) in the three groups was tested by enzyme linked immunosorbent assay (ELISA).@*RESULTS@#Immunocytochemical staining showed positive vimentin expression in NF and TSF. Results also indicated the weak positive expression of α-smooth muscle actin (SMA) and fibroblast activation protein (FAP) in TSFs and the negative expression of α-SMA and FAP in NFs. MTT assay showed that cell proliferation in the TSF-SPAC group was significantly different from that in the NF-SPAC and SPAC groups (P<0.05). Cell proliferation was not different between the NF-SPAC and SPAC groups (P>0.05). Transwell and scratch assays showed no difference in cell invasion and migration among the groups (P>0.05). ELISA showed that no significant difference in VEGF expression among the three groups (P>0.05).@*CONCLUSIONS@#TSFs may be involved in SPA biological behavior by promoting the proliferation of SPACs but has no effect on the invasion and migration of SPACs in vitro. Hence, TSF may be a new therapeutic target in SPA treatment.
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Humans , Adenoma, Pleomorphic/metabolism , Vascular Endothelial Growth Factor A , Culture Media, Conditioned/metabolism , Fibroblasts/metabolism , Salivary Glands/metabolismABSTRACT
Objective:To investigate the effects of exenatide on podocyte in diabetic nephropathy mice.Methods:Diabetic nephropathy mice models were induced by using streptozocin-treated C57BL/6J mice on high fat diets, which were randomized by random number table to diabetic nephropathy control group (DN group, n=8) and exenatide treatment diabetic nephropathy group (DN+ Ex group, n=8). The C57BL/6J mice on normal chow diet were used as normal control group (NC group, n=8). After the intervention, blood glucose, renal function, and urine albumin/creatinine ratio were measured. Pathological glomerular changes were observed by pexiodic acid-Schiff stain (PAS) staining. The mRNA expression of profibrotic molecules Collagen Ⅳ, transforming growth factor-β (TGF-β), and Fibronectin in glomerular lysates were measured by realtime quantitative PCR (RT-PCR). Podocyte injury and apoptosis were evaluated by immunofluorescent staining and transmission electron microscopy. The expression of Nephrin, Cleaved caspase-3, protein kinase B (Akt), and phosphorylated Akt (p-Akt) in glomerular lysates were examined by western blotting. Results:Compared with the DN group, urine albumin/creatinine ratio was significantly decreased in the DN+ Ex group ( P<0.01). PAS staining and analysis found that exenatide administration ameliorated mesangial matrix expansion and glomerular hypertrophyin in DN group ( P<0.05). RT-PCR analyses showed that the glomerular expression for Fibronectin, TGF-β, and Collagen Ⅳ were significantly decreased in the DN+ Ex group compared with the DN group ( P<0.01). Immunofluorescent staining and transmission electron microscopy revealed that exenatide treatment improved podocyte injury and apoptosis in DN group. Western blotting analyses showed that exenatide increased the Nephrin expression, decreased the Cleaved caspase-3 expression, increased the p-Akt expression in glomerular lysatesin diabetic nephropathy mice ( P<0.01). Conclusion:Exenatide attenuates podocyte injury and apoptosis and proteinuria, and prevents the progression of diabetic nephropathy mice. Phosphatidylinositol 3-kinase (PI3K)/Akt pathway in glomerular lysates may be related to the protective effects of exenatide.
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Objective To study the effect of vibration training with adjusted frequency on changes in explosive force of lower limbs, balance and muscle function around ankle joints of patients with functional ankle instability (FAI), so as to provide an empirical basis for rehabilitation training of FAI patients in clinic. Methods Twenty-six FAI patients were randomly divided into the experimental group (n=14) and the control group (n=12). The experimental group received 8-week rehabilitation training with vibration intervention, while the control group only received 8-week rehabilitation training. Changes in maximum power, average power, maximum speed and average speed of the injured limb during vertical jump with single leg, changes in distances during long jump, changes in time during one-leg standing with eyes open and closed，changes in contract time (tc), relax time (tr) and displacement (Dm) of medial gastrocnemius (GM), lateral gastrocnemius (GL) and tibialis anterior (TA) muscles before and after training were measured and compared. Results In the experimental group, the maximum power and maximum speed of the injured limb during vertical jump with single leg, the distance during long jump with single leg and the time during one-leg standing with open and closed eyes were significantly improved, and the increase was higher than that of the control group. The increase of tc of all muscles in the experimental group was smaller than that of the control group, but tr and Dm did not show any regularity. Conclusions Vibration training with adjusted frequency can effectively improve the explosive force and balance ability of lower limbs of FAI patients, and promote the tc shortening of GL, GM and TA muscles, but whether vibration training with adjusted frequency can reduce muscle tension and promote muscle relaxation is still not clear.
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Objective To investigate the effects of lentivirus-mediated RNA interference (RNAi) targeting DNA binding protein A (dbpA) on the proliferation and the biological behavior of colorectal cancer cell line SW620.Methods The experiment was divided into 3 groups:KD group (siRNA-dbpA,lentivirus interference group),CON group (non-specific sequence group) and NC group (blank control group).The lentiviral vector siRNA-dbpA was constructed and verified by PCR and DNA sequencing.SW620 cells were transfected with siRNA-dbpA plasmid,nontargeting siRNA plasmid,or empty plasmid.After 48 h the transfection,the cells were examined for dbpA expression using Western blot.After 72 hrs transfection,flow cytometry was used to detect the cell apoptosis and cell cycle changes.The cell growth inhibition rate was detected by MTT (4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide) assay,and then clone formation was detected,and the ability of SW620 cells to form tumors in vivo after dbpA was silenced was studied in nude mice.Results PCR analysis and DNA sequencing demonstrated that the RNAi sequence targeting dbpA gene was successfully inserted into the lentiviral vector.siRNA-dbpA transfection resulted in reduced expression of dbpA in SW620 cells.After transfection,the apoptosis rate of siRNA-dbpA-transfected cells increased to 26.60％ ± 0.38％,significantly higher than that in cells transfected with the nontargeting plasmid or the empty plasmid 12.54％ ± 0.25％ and 4.46％ ± 0.19％,respectively (F =28.159,P ＜0.01).The growth inhibition test indicate that the OD value of the fifth day in siRNA-dbpA group was 0.194 ±0.037,significantly lower than that in the other two groups 0.814 ±0.043 and 1.625 ±0.061,respectively(F =23.214,P ＜ 0.01).The colony formation number is 37 ± 3,64 ± 5and 175 ± 10 respectively,siRNA-dbpA is significantly higher than that in the other two groups(F =40.254,P ＜ 0.01).After the completion of nude mouse transplantation tumor model,through the detection of tumor volume,KD group (group siRNA-dbpA) tumor volume after 14 d and CON and NC group had obvious difference (F =38.256,P ＜ 0.05),and after 21d is more significant difference in tumor size (F =40.241,P ＜ 0.01),can be clearly observed after 35 d KD group (group siRNA-dbpA) growing tumors had differences with the control group (F =30.257,P ＜ 0.05).Conclusion Lentivirus-mediated RNAi targeting dbpA can effectively suppress the expression of dbpA in colorectal tumor in nude mice,it is proved that dbpA silencing has a significant inhibitory effect on the growth of living tumor cells and decrease the proliferation of the colorectal cells.
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Objective@#To investigate the clinical manifestations and pathological changes of benign lymphoadenosis of oral mucosa.@*Methods@#The clinical data of 98 cases of benign lymphoadenosis of oral mucosa were analyzed.@*Results@#The clinical manifestations of benign lymphoadenosis of oral mucosa included erosive ulcer (64%) and nodule (9%) and the rate of misdiagnosis was 98%. Neutrophil infiltration occurred in the epithelium of 51% cases and the lymphocyte was diffusely infiltrated in lamina propria of 83% cases.@*Conclusions@#When the mucous membrane of the lamina propria is characterized by complex cell components, diffuse infiltrating lymphocytes and infiltration of neutrophils in mucosal epithelium without erosion and ulceration, it is necessary to highly suspect benign lymphoadenosis of oral mucosa. Finding the focal aggregation of lymphoid follicles or lymphocytes is helpful for the correct diagnosis.
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Objective: To compare the sedation induced by target-controlled infusion of propofol with that by propofol-remifentanil in third molar exaction surgery. Methods: 60 patients for third molar exaction were divided randomly into 2 groups(n = 30): group P(propofol group) and group PR(propofol-remifentanil group). In group P,a titrated infusion of propofol was started until the OAA/S score had reached level 3 in the patients,then the surgery began. In group PR,a infusion of remifentanil with a target plasma concentration of 1 ng /ml and a titrated infusion of propofol was started until the OAA/S score had reached level 3,then the surgery began. In all patients,the heart rate,blood pressure,respiratory rate,oxyhemoglobin saturation and narcotrend index were recorded during the operation. The reactions of the patients in the operation were recorded. The satisfaction of the patients and surgeons was asked. Results: The respiratory rate and the oxyhemoglobin saturation in group PR was lower than those in group P(P ＜ 0. 05). No obvious adverse reaction was observed in the 2 groups. The satisfaction of the patients in the 2 groups was 30 /30 and 30 /30(P＞ 0. 05). Conclusion: The sedation induced by target-controlled infusion of propofol or propofol-remifentanil in third molar extraction is safe. The sedation under target-controlled infusion of propofol-remifentanil is better than that by propofol when inhalating oxygen.
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Objective To investigate the effect of postoperative analgesia by using loxoprofen sodium on dental implant patients at different time points. Methods A total of 400 patients with dental implant treatment were divided into two groups by random number table method. The experimental group was firstly given loxoprofen sodium tablets (60 mg) in 30 minutes preoperatively, and the control group was firstly given on three hours after surgery (60 mg). Local anesthesia was used to all dental implant surgery. Using the Wong-Baker Smile Assessment method in operation and Numerical Rating Scale (NRS) in postoperative respectively to assess pain level in surgery and 3 h, 6 h, 12 h and 24 h after surgery. Results The percentages of painless patients of the experimental group and the control group in operation were 99%(198/200) and 97%(194/200), and there was no significant difference between them (χ2=2.041, P>0.05); the percentages of painless patients of the experimental group at 3 h, 6 h, 12 h after surgery were 60.5%(121/200), 79.0%(158/200), 83.5%(167/200), and the control group were 47.0%(94/200), 64.5%(129/200), 71.5%(143/200), and there was significant difference between the control group and the experimental group (χ2=14.255,15.447, 11.165, P=0.007, 0.004, 0.011); however, in the two groups, there was no significant difference at 24 h after surgery, the experimental group was 93.0% (186/200), the control group was 89.5% (179/200) (χ2=2.468, P>0.05). Conclusions Preoperative administration with loxoprofen sodium tablets can significantly reduce the risk of postoperative pain, and could be used as a conventional implant surgery analgesic program.
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Objective The aim of this study is to compare sedation titrated under target-controlled infusion of propofol and propofol-remifentanil for impacted supernumerary teeth extraction surgery for children. Methods A total of 60 children with anterior maxillary region impacted supernumerary teeth extraction surgery were divided randomly into two groups, namely, propofol group (group P, n=30) and propofol-remifentanil group (group PR, n=30). In group P, a titrated infusion of propofol was started until the modified observer's assessment of alertness/sedation (OAA/S) scale reached level 3 before the actual surgery. In group PR, a remifentanil infusion with a target plasma concentration of 1 ng·mL⁻¹ was started until the operation was finished. A titrated infusion of propofol was also started until the modified OAA/S score reached level 3 before the actual surgery. The Houpt behavior scale was adopted to evaluate the cooperation of each patient in both groups. The heart rate, blood pressure, respiratory rate, oxyhemoglobin saturation, and Narcotrend index, complications, adverse reactions and propofol infusion of all patients were recorded during the operation. Results The Houpt behavior scales in group PR were better than those in group P (P<0.05). The oxyhemoglobin saturation and respiratory rate in group PR were lower than that in group P (P<0.05). The heart rate, blood pressure and NI in two groups were no significant difference (P>0.05). The incidence of respiratory depression and anterograde amnesia in group PR were higher than that in group P (P<0.05). Conclusion Sedation titrated under the target-controlled infusion of propofol and that titrated under propofol-remifentanil for impacted supernumerary teeth extraction surgery for children are safe. The sedation titrated under target-controlled infusion of propofol-remifentanil is better than sedation by propofol when inhaling oxygen.
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Objective To monitor and analyze the antigenicity of Streptococcus pneumonia polysac-charide, its derivatives and conjugates by three immunological assays .Methods Inhibition ELISA and rate nephelometry(RN) were established for this study.Antigenicity of serotype 23F pneumococcal conjugates and their intermediates were analyzed by double immunodiffusion assay , inhibition ELISA and RN .The re-sults derived from three assays were comparatively analyzed to evaluate the changes of antigenicity during the preparing process of serotype 23F conjugate.Results Double immunodiffusion assay, inhibition ELISA and RN were all applicable to antigenicity analysis during the process of conjugate preparation .Inhibition ELISA could quantitatively detect a slight difference of polysaccharide antigenicity during the preparing process . Conclusion The antigenicity of polysaccharide during the preparing process of pneumococcal conjugates could be analytically monitored by using three immunological assays .This study provided evidence for suc-cessfully using immunological assays as the quality control means during the preparing process of pneumococ -cal conjugates .
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AIM: To observe the changes of morphology, the activity of myeloperoxidase (MPO) and membrane pump activities of spleen tissue in acute renal failure (ARF) rabbits, and to inquire into the role of spleen on pathogenesis of immune function disorders during ARF. METHODS: 42 rabbits were divided into control group, HgCl_2 group and glycerinum group. The ARF model was established by hypodermic injection of 1% HgCl_2 at dose of 1.3 mL/kg in HgCl_2 group, intramuscularly injection of 50% glycerinum at dose of 10 mL/kg in glycerinum group, respectively, and the animals were divided into the 12 h, 24 h, 48 h secondary groups (6 rabbits each group). At different time points, the rabbits were cannulated to facilitate the collection of blood sample to examine the biochemical indexes of renal function. The spleen microscopic sections were prepared for observing the morphology. The spleen homogenate was made for determining the activities of MPO and membrane pumping. RESULTS: Pathological sections of spleen showed that the different degree of congestion was found and spleen trabecula was increased in two model groups at multiple-time points. The MPO activity of spleen homogenate in HgCl_2 group and glycerinum group at all time points were obviously higher than that in control group, and at 24 h, the MPO activitie in two model groups was significantly increased than that in the same group at 12 h and 48 h. The activities of Na~+-K~+-ATPase, Ca~(2+)-ATPase, Mg~(2+)-ATPase, Ca~(2+)-Mg~(2+)-ATPase of spleen homogenate in two model groups at multiple time points were significantly lower than those in control group. Following ARF development, the ATPase activitie in two model groups at 48 h was lower than that at 12 h except the Mg~(2+)-ATPase in glycerinum group. CONCLUSION: Spleen as an immune organ has histological damage, arrest of polymorphonuclear neutrophils and dysfunction of membrane pump during the development of ARF in rabbits, leading to immune disorders.
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The treatment for chylothorax depends on the underlying causes and the individual clinical cir-cumstances.It is difficult to select the best treatment measure.The main options include medium-chain tri-glyceride diet,total parenteral nutrition,drug therapy,thoracentcsis,pleurodesis,pleuroperitoneal shunting,embolization or ligation of the thoracic duct.This paper provides a brief review of the treatment for chylotho-rax.