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Objective:To investigate the number of necroptotic renal tubular epithelial cells in renal tissues of patients with chronic kidney disease (CKD) and the correlation with clinicopathologic parameters, and explore its role in the progression of the excessive loss of renal tubular cells and chronic kidney injury.Methods:Renal tissue samples from 60 patients (18-65 years old) with CKD proven by kidney biopsy in the First Affiliated Hospital of Hainan Medical University from June 2017 to June 2019 were collected. According to internationally accepted K/DOQI guidelines, the patients were divided into 1-4 stages of CKD, with 15 cases in each stage. The number of necroptotic renal tubular epithelial cells in patients with different stages of CKD was detected using receptor-interacting protein 3 (RIP3) and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) fluorescent staining, and the expression of RIP3 and MLKL, marker protein of necroptosis, was detected by immunohistochemistry. Pearson correlation analysis was used to analyze the correlation between the percentage of necroptotic renal tubular epithelial cells and clinicopathologic parameters. In addition, the expression of angiotensinogen Ⅱ receptor (AT2R) in renal tissue and its correlation with the percentage of necroptotic renal tubular epithelial cells were analyzed.Results:With the development of CKD, the structural destruction of renal tubules in patients with CKD was gradually aggravated, and the renal tubules in the corresponding areas were atrophied, accompanied by worsening interstitial fibrosis. The adjacent renal tubules were focally dilated and numerous protein tubules were seen in the tubules. Importantly, renal tubular injury score in second and third stage of CKD was significantly higher than that in control group (both P<0.01). TUNEL+RIP3 immunofluorescence staining results showed that the percentage of TUNEL/RIP3 double positive renal tubular epithelial cells (necroptotic renal tubular epithelial cells) in renal tubules of the second and third stage of CKD was higher (all P<0.01). Immunohistochemical results showed that RIP3, MLKL and AT2R proteins were mainly expressed in cytoplasm of renal tubular epithelial cells, and the expression of RIP3, MLKL and AT2R in renal tubular epithelial cells was higher in the second and third stage of CKD patients (all P<0.05). Pearson correlation analysis showed that the percentage of necroptotic renal tubular epithelial cells was positively correlated with blood urea nitrogen ( r=0.514, P=0.003), serum creatinine ( r=0.507, P=0.019), serum cystatin C ( r=0.571, P=0.026), serum uric acid ( r=0.592, P=0.008), renal tubules injury score ( r=0.901, P<0.001), renal interstitial fibrosis index ( r=0.700, P=0.001) and the expression of AT2R protein in renal tissue ( r=0.715, P=0.001). Conclusions:As CKD progresses, necroptosis of renal tubular epithelial cells in CKD patients occurs. The necroptotic cell death may be an important factor leading to renal tubular epithelial cell excessive death and the progression of chronic kidney injury. Furthermore, necroptosis of renal tubular epithelial cells may be related to the high expression of AT2R in kidney tissue.
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Objective To evaluate the prevalence of chronic kidney disease (CKD) in Chinese adult health check-up population,and to compare with the prevalence of CKD in the study of the general population as well as the large CKD cross-sectional study in China.Methods Epidemiological studies about CKD in Chinese adults health check-up population from January 2007 to December 2017 were searched in PubMed,SinoMed,CNKI,VIP and Wanfang Data.Meta-analysis of the prevalence of CKD was performed with software of Stata 12.0.Subgroup analyses of CKD staging,urban and rural,as well as geographical areas of the general population were executed.Results Twenty-two studies from adult health check-up population were included (238 349 persons).Egger's regression showed no publication bias (P ＞ 0.05).The unstandardized prevalence rate of CKD was 12.49％ (male 12.8％,female 12.5％).The respective unstandardized prevalences of proteinuria,hematuria and eGFR decline were 5.90％,5.83％ and 2.75％.The unstandardized prevalences of CKD in urban and rural population were 13.21％ and 11.90％.The stages of CKD were mainly concentrated in the early stages.There was no significant difference in the non-standard detection rate of total eGFR decline among the adult medical examination population,the general population and the population studied cross-sectionally (P ＞ 0.05).Furthermore,no significant difference in the non-standard detection rate of total hematuria and male hematuria was found between the adult health check-up population and the general population.In addition,the total proteinuric non-standard detection rate of the adult general population was similar with that of population studied cross-sectionally (P ＞ 0.05).Conclusions The prevalence of CKD in Chinese adults is higher,the overall prevalence is however underestimated.The results of epidemiological investigation in adult health check-up population are similar to those of the general population,especially in men.
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Objective To investigate the differences in the biochemical and pathological features and therapeutic response between the elderly-onset autoimmune hepatitis (AIH) and young-onset AIH.Methods From October 2013 to November 2016,a total of 87 consecutive AIH patients diagnosed by liver biopsy were enrolled and divided into elderly-onset group (≥60 years) and the young-onset group (＜ 60 years).The biochemical and pathological features and therapeutic response of the two groups were compared.Mann Whitney test and chi square test were performed for statistical analysis.Results As AIH diagnosed,the proportion of liver cirrhosis detected by imaging examination of elderly-onset group and the young-onset group were 67.7％(21/31) and 35.7％(20/56),respectively,the former was higher than the latter,and the difference was statistically significant (X2 =8.214,P=0.004).The proportion of patients complicated with extrahepatic autoimmune diseases of the elderly-onset group was 9.7％ (3/31),which was lower than that of the young-onset group (28.6％,16/56),and the difference was statistically significant (X2 =4.173,P=0.041).The median levels of alanine aminotransferase (ALT) and albumin of the elderly-onset group were 50.0 U/L (22.0 U/L,193.0 U/L) and 34.8 g/L (31.3 g/L,40.5 g/L),which were lower than those of the young-onset group (146.0 U/L (43.8 U/L,390.5 U/L),39.4 g/L (35.8 g/L,44.6 g/L),and the differences were statistically significant (Z=-2.109,-2.092;both P＜ 0.05).Between the two groups,there was no statistically significant difference in inflammation grade,fibrosis stage,interface hepatitis,lymphoplasmacytic infiltration,positive rate of hepatocyte rosette pattern,positive rate of centrilobular necrosis and small bile duct injury (all P＞0.05).After treatment,among 17 patients of elderly-onset group,seven patients obtained biochemical remission and the median time to remission was 3.2 months;among 37 patients of young-onset group,18 patients (48.6％) obtained biochemical remission and the median time to remission was 2.3 months.There was no significant difference in the percentage of patients achieved biochemical remission and the median time between the two groups (both P＞0.05).Conclusion The percentage of complicated with extrahepatic autoimmune diseases of elderly-onset group is lower than that of young-onset group,which indicates that age-associated immune dysfunction may involve in the genesis and development of elderly-onset AIH.
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AIM: To explore whether tumor necrosis factor-α (TNF-α) induces necroptosis in murine long bone osteocyte-like cell line MLO-Y4 and the possible mechanism.METHODS: The MLO-Y4 cells were divided into control group, TNF-α group, TNF-α+necrostatin-1 (Nec-1) group, TNF-α+Z-VAD group and TNF-α+receptor-interacting protein 3 (RIP3)-siRNA group.The death rate of MLO-Y4 cells was assessed by flow cytometry with Annexin V-FITC/PI staining.The morphological features of the cells were observed under transmission electron microscope (TEM).The protein levels of RIP1, RIP3 and cleaved caspase-3 were determined by Western blot.Finally, the numbers of total cells and RIP1-RIP3-positive cells were observed under laser scanning confocal microscope.The production of reactive oxygen species (ROS) in the cells was measured by DCFH-DA staining.RESULTS: Compared with control group, the apoptotic or necroptotic rate of the cells induced by TNF-α was increased significantly (P<0.01).The increased apoptotic or necroptotic rate was dramatically reduced by treating with Nec-1, Z-VAD or RIP3-siRNA transfection (P<0.01).In TNF-α group and TNF-α+Z-VAD group, a lot of MLO-Y4 cells with typical necroptotic morphological features were observed under TEM.However, obvious necroptotic cells were not found in Nec-1 or RIP3-siRNA treatment group.The protein level of RIP1 in the cells treated with Nec-1 was sharply lower than that in TNF-α group (P<0.01).However, Z-VAD did not reduce the elevated levels of RIP1 and RIP3.RIP3-siRNA effectively down-regulated the protein level of RIP3 compared with TNF-α group (P<0.01).Nec-1 effectively down-regulated the protein levels of RIP1 colocalized with RIP3 compared with TNF-α group (P<0.01).However, Z-VAD did not reduce the levels of RIP1 colocalized with RIP3.Nec-1, Z-VAD and RIP3 siRNA significantly decreased the ROS levels (P<0.01).CONCLUSION: TNF-α induces the necroptosis of MLO-Y4 cells.RIP3 play vital roles in the cell necroptotic signal pathway.ROS may be the executor of necroptosis of MLO-Y4 cells.
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This study aimed at exploring a new method--microtox technology to evaluate the comprehensive toxicity of safflower injections from different drug manufacturers.In the experiment,we investigated the characteristic parameters of vibrio fischeri (CS234) under different conditions,and then locked the optimum parameters for safflower injection assay:recovery liquid volume 0.9 mL in freeze-dried vial,50 μL bacteria suspension in each sample,and the detection time of 10 mins after adding the bacteria suspension.Under this optimum detection condition,we found that the EC50 values were 3.36％,5.58％ and 4.33％,being significantly different between 3 representative drug manufacturers (P ＜ 0.05).In conclusion,it was implied that the biological detection standards of safflower injections need improving,while the microtox-based fast test of the comprehensive toxicity of safflower injection showed a favorable and prospective application to controlling the product qualities of different manufacturers.
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This study aimed at exploring the application of microtox technology to the evaluation of comprehensive toxicity of Sheng Mai injections.Characteristic parameters of vibrio fischeri (CS234) were measured by methodology inspection under different conditions to optimize the reaction condition with reliable technology.It was the first experiment to accomplish the comprehensive toxicity of Sheng Mai injections from various manufacturers using vibrio fischeri under the target condition.It was found that parameters of the optimum condition contained 0.9 mL recovery liquid volume in each freeze-dried vial and 50 μ L bacteria suspension of each sample,being included in 2 mL solution in aggregate under 10 mins' detection time after adding bacteria suspension with the favorable pH value ranging from 5 to 10 and luminous intensity from 0.8 to 1.2 million within 10 mins.The relative standard deviation values of replication experiment and precision test were all below 15％.Under this optimum detection condition,it was found that the EC50 values were 22.10％,34.10％ and 46.04％,respectively,presenting significant statistical differences (P＜0.05).These results demonstrated that the growth of biological detection standards of Sheng Mai injection was highlighted a long way to go.Besides,the microtox-based fast test for the evaluation of the comprehensive toxicity of Sheng Mai injection showed a prospective application to controlling the quality of different products from manufacturers.
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[ ABSTRACT] AIM: To explore whether autophagy is involved in the excessive death of renal tubular epithelial cells in subtotal nephrectomy ( SNx) rats and the relationship between autophagy and necroptosis in the kidney of SNx rats. METHODS:Male Sprague-Dawley rats were randomly assigned to control group ( n=6 ) and SNx group ( n=42 ) .The rats in SNx group were subjected to SNx.Sham surgery was performed in the rats in control group.The rats in SNx group were divided into subgroups at 0, 4, 8 and 12 weeks ( n=6) and the other rats in SNx group were divided into SNx+vehi-cle group, SNx+necrostatin-1 (Nec-1) group and SNx+3-methyladenine (3-MA) group.The expression of RIP1, RIP3, LC3 and beclin-1 at mRNA and protein levels was measured at 0, 4, 8 and 12 weeks by qPCR and immunohistochemistry. The effects of Nec-1 or 3-MA on the protein expression of LC3-I, LC3-II and beclin-1, and production of reactive oxygen species ( ROS) in the rat kidney were determined by Western blot and DCFH-DA staining.The death of renal tubular epi-thelial cells in the SNx rats was observed by TUNEL staining and electron microscopy.Finally, the effects of Nec-1 and 3-MA on blood urea nitrogen ( BUN) , serum creatinine ( SCr) and the pathological changes of the renal tissues were ana-lyzed.RESULTS:The highest mRNA and protein levels of RIP1, RIP3, LC3 and beclin-1 appeared at the 8th week after SNx (P cells were decreased in the SNx rats treated with Nec-1 and 3-MA (P<0.01), but 3-MA did not reduce the increased con-centration of ROS.In addition, treatment with Nec-1 and 3-MA obviously reduced BUN, SCr (P<0.05), glomeruloscle-rosis index and tubulointerstitial injury score (P<0.01).CONCLUSION:Autophagy participates in the excessive death of renal tubular epithelial cells in SNx rats.Inhibition of autograph prevents necroptotic cell death of renal tubular cells, and alleviates chronic renal injury in SNx rats.
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Objective To compare the clinical effects of extensive or limited arthroscopic debridement in varying degrees of knees osteo-arthritis. Methods 32 cases with knees osteoarthritis who underwent extensive debridement under arthroscopy were selected as the observa-tion group, and 30 cases with knees osteoarthritis who underwent limited debridement were selected as the control group. The excellent rate of joint function, Lysholm and VAS scores were compared between the two groups. Results The excellent rate of joint function were 90. 0%and 91. 4% in observation group and control group respectively, without significant difference (P>0. 05). Lysholm scores of patients with grade Ⅱ osteoarthritis was much higher in observation group than that in control group after operation (P0. 05). No statistically significant difference of VAS scores on 1 year after operation were found (P>0. 05). Conclusion Extensive debridement and limited debridement under arthroscopy have similar effects for knees osteoarthritis. Extensive arthroscopic debridement is an ideal treatment method for Ⅱ grade of knees osteoarthritis.
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<p><b>OBJECTIVE</b>To evaluate the efficacy and safety between endoscopic and microscopic surgery for transsphenoidal pituitary adenoma.</p><p><b>METHODS</b>Randomized or semi-random controlled trials comparing endoscopic with microscopic surgery for transsphenoidal pituitary adenoma and published between January 2000 and July 2013 were recruited. This meta-analysis (RevMan 5.1 software) was conducted to estimate short-term and long-term complications. Fixed random effect model or semi-random effect model was established to analyse the data.</p><p><b>RESULTS</b>Twelve randomized or semi-random controlled trials were included in this study. Among 848 patients studied, 380 of them were treated with endoscopic and 468 were treated with microscopic. The analysis of the basic characteristics of these patients included in these studies showed that: compared with microscopic, the follow-up of patients in endoscopic group was shorter [OR = -2.29, 95%CI (-4.18, -0.39), P = 0.02], while there were no significant difference in gender proportion and the age of patients between the two groups (P > 0.05) . Also, compared with endoscopic, there was a significant addition in the incidence of diabetes insipidus [OR = 0.45, 95%CI(0.30,0.66), P < 0.0001] and other complications [OR = 0.26, 95%CI ( 0.12, 0.57), P = 0.0008] in the microscopic group, the blood loss during surgery was more [OR = -0.62, 95%CI(-1.19, 0.05), P = 0.03], the rate of complete tumor resection lower [OR = 0.61, 95%CI(0.39, 0.96), P = 0.03], and the hospitalization [OR = -1.53, 95%CI(-2.18, -0.88), P < 0.00001] was also significantly longer, there was no significant difference in the incidence of cerebrospinal fluid leakage, operation time and vision improvement rate (P > 0.05).</p><p><b>CONCLUSION</b>For patients with pituitary tumors, endoscopic surgery may be more suitable compared with microscopic.</p>
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Humanos , Adenoma , Endoscopia , Medicina Baseada em Evidências , Microcirurgia , Procedimentos Neurocirúrgicos , Métodos , Neoplasias Hipofisárias , Cirurgia Geral , Ensaios Clínicos Controlados Aleatórios como Assunto , Resultado do TratamentoRESUMO
BACKGROUND:Bone marrow mesenchymal stem cells(BMSCs)are widely utilized as seed cells or carriers in bone tissue engineering and gene therapy.Thus,how to obtain BMSCs with high purity arose more attentions of researchers.OBJECTIVE:To observe the effects of different separation methods and culture conditions on biological characteristics of rabbit BMSCs.METHODS:BMSCs were obtained by whole bone marrow culture,density grand centrifugal and red blood cell lysis.At 48 hours after culture,the cell numbers were counted,the time of passage was recorded,in addition,the cell morphology was observed by phase contrast microscope,and the CD44 antigen expression was identified using flow cytometry.The 3rd and the 7th generation aging cells were cultured with DMEM-LG,MEM-HG,and DMEM/F12 culture medium.MTT and count cell plat were used to evaluate the growth of BMSCs.Phase contrast microscope was used to observe the morphological changes of aging cells.RESULTS AND CONCLUSION:BMSCs could be separated by each method.The adherent cells showed shuttle or multiple angle shapes,with rich cytoplasm,and positive for CD44 antigen.The more cell number and shortest primary culture time was presented in red blood cell lysis group(P ＜ 0.05).DMEM/F12 could promote the proliferation of quiescent cells.And the cells prevented the better viability.The method of read blood cell lysis improving the efficiency of BMSCs adherent is an effective method of extraction of BMSCs.DMEM/F12 could promote the proliferation Maybe,DMEM/F12 is more suitable for BMSCs.
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Objective To establish a stable model of rat orthotopic left lung transplantation using direct suture of vessels and bronchi. Methods Ten Sprague Dawley (SD) rats weighted 250 to 350 g were used as lung donors and recipients respectively. Airway and pulmonary vessels were reconstructed microsurgically using continuous running suture technique. Survival time were recorded and donor lungs were checked by autopsy. Results All 10 rats received left lung transplantation were weaned from ventilator successfully. Both of cold ischemia time and warm ischemia time were about 40 minutes. The total procedure took about 130 minutes. Autopsy was used to check the patency of anastomotic sites. No thrombosis or air leak was found. Conclusions Direct microsurgical surture can be used to establish an experimental model of orthotopic left allograft lung transplantation in rats. This method is proved to be stable, reliable and similar to clinical practices.
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OBJECTIVE@#To provide the anatomic data for operation on the middle ear through the observation and measurement of related anatomic structure.@*METHOD@#Forty human temporal bones of 20 voluntary bone donors were dissected, relative anatomical data of operation were observed and measured under operating microscope through posterior tympanum approach entering posterior tympanum.@*RESULT@#The average distances from suprameatal spine to short crus of incus, pyramid segment of facial canal were 19.14 mm, 16.30 mm, respectively. The average distances from pyramid segment of facial canal to the surface of mastoid, crotch of chorda tympani nerve, posterior wall of auditory meatus were 20.84 mm, 11.28 mm, 4.40 mm, respectively. The average length of facial nerve in the horizontal segment, the perpendicular paragraph was about 11.60 mm, 15.30 mm, respectively. The average distance from pyramidal eminence to the anterior lip of round window niche, from oval window to round window niche, from incudostapedial joint to round window niche was 4.46 mm, 3.74 mm, 3.80 mm, respectively. The included angle of facial nerve in the horizontal segment and chorda tympani nerve with facial nerve in the perpendicular paragraph was 110.4 degrees, 24.8 degrees, respectively. Horizontal semicircular canal and facial nerve in the level paragraph was 17.5 degrees, long process of incus and incus buttress was 46.0 degrees.@*CONCLUSION@#The position of anatomic structure in middle ear was constant and the relationship including distance and angle between anatomic structures changed in limited region. The anatomical parameters provide a reference value for avoiding the injury during the operation.
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Adulto , Feminino , Humanos , Masculino , Nervo da Corda do Tímpano , Orelha Média , Cirurgia Geral , Nervo Facial , Janela da Cóclea , Cirurgia Geral , Osso TemporalRESUMO
Objective To evaluate the clinical effect of a combination application of meglumine adenosine cyclophosphate and metoprolol in chronic heart failure.Methods Put 50 patients divided into two group randomly:group A(treated group) which has 26 cases and group B(control group) which has 24 cases.Patients of group B will receive a basal treatment in a routine way while patients of group A will receive a basal treatment as well as a additional treatment of meglumine adenosine cyclophosphate and Metoprolol.The clinical effect and changes of heart function will be evaluated in the two group with different treatment after three weeks,left ventricular ejection fraction and left nentricular volume at end-diastole,left nentricular volume at end-systolic were measured by M-mode.Echocardiography before and after curative in two group.Results Patients in the group with a combination application of meglumine adenosine cyclophosphate and metoprolol has a better clinical effect and a improved heart function than the control group.Conclusion A combination application of meglumine adenosine cyclophosphate and metoprolol has good curative effect in treating heart failure.