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Objective:To investigate the feasibility of anthocyanins(C3G)antioxidant inhibition of autophagy to al-leviate epilepsy.Methods:Seventy-five SD rats were randomly divided into 5 groups:Control group,pentetrazole(PTZ)group,hydrogen peroxide(H2O2)intervention group,3-methyladenine(3-MA)intervention group,and C3G intervention group.The seizure grade,latency,and frequency were documented.Electroencephalography was employed to detect abnormal electrical discharges in the brain across.Patch clamp technique was utilized to measure action poten-tials in hippocampal neurons for each group.The concentration of 4-hydroxynonenoic(4-HNE)hippocampus was deter-mined using a specific kit.Ultrastructural alterations in hippocampal neurons were examined through electron microsco-pyissl staining was performed to assess neuronal damage within the hippocampus.Immunohistochemical staining and Western Blot were conducted to evaluate expression levels of 15-LOX,GPX4,and LC3 proteins within the hippocampus of rats.Results:Compared with the control group,the PTZ group was completely ignited and the modeling was success-ful.Compared with other epilepsy groups,the seizure grade of C3G group decreased,abnormal discharge decreased,latency increased,hippocampal neuron excitability decreased,nishi content increased,4-HNE content,15-LOX expression and LC3Ⅱ/LC3Ⅰ ratio decreased,but GPX4 expression increased(P＜0.05).Conclusion:The oxidative stress induced by epilepsy can induce excessive autophagy of neurons,and C3G can alleviate the occurrence and devel-opment of epilepsy by anti-oxidation and inhibition of autophagy.
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OBJECTIVE@#To investigate the effect of silencing DNA methyltransferase 1(DNMT1) to the methylation of the promoter of the tumor suppressor gene wnt-1 (WIF-1) in human chronic myeloid leukemia (CML) cells.@*METHODS@#DNMT1 siRNAi plasmid was constructed and DNMT1 siRNAi was transfected into CML K562 cells. RT-PCR and Western blot were used to detect the expression of DNMT1 gene and related protein, and methylation PCR was used to detect WIF-1 gene promoter methylation level. The trypan blue exclusion and MTT assay were used to detect the cell proliferation, flow cytometry were used to detect the cell apoptosis rate, colony formation assay was used to detect cell colony formation ability. Expression of Wnt/β- catenin and its downstream signaling pathway proteins were detected by Western blot after DNMT1 gene was silenced.@*RESULTS@#The expression level of DNMT1 mRNA and its related protein in the experimental group were significantly lower than those in the control group and negative control group (P<0.05). After 72 hours of successful transfection, the WIF-1 gene in the control group and negative control group were completely methylated, while in the experimental group, the methylation level significantly decreased. The results of MSP showed that the PCR product amplified by the unmethylated WIF-1 primer in the experimental group increased significantly,while by the methylated WIF-1 primer decreased significantly. After 48 h of transfection, the OD value, viable cell number and colony formation of the cells in experimental group were significantly lower than those in the negative control group and the control group (P<0.05). The apoptosis rate of the cells in experimental group was significantly higher than those in the negative control group and control group (P<0.05). The expression levels of β- actin, myc, cyclin D1 and TCF-1 in K562 cells in the experimental group were significantly lower than those in the negative control group and control group (P<0.05).@*CONCLUSION@#Silencing DNMT1 gene can inhibit the proliferation and promote the apoptosis of K562 cells. The mechanism may be related to reverse the hypermethylation level of the WIF-1 gene promoter, thereby inhibit the activity of the Wnt/β- catenin signaling pathway.
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Humans , Adaptor Proteins, Signal Transducing/metabolism , DNA Methylation , K562 Cells , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/genetics , Repressor Proteins/metabolismABSTRACT
Objective:To investigate the neuroprotective effect of ibuprofen, and influence of ibuprofen in hippocampal nod-like receptor protein 3 (NLRP3) inflammatome and its related products in chronic epilepsy rats models.Methods:Thirty male SD rats were randomly divided into 3 groups: control group, pentylenetetrazol (PTZ) group and PTZ+ibuprofen group ( n=10). Rats in the PTZ group were intraperitoneally injected with PTZ (35 mg/kg) once every one d, and rats in the PTZ+ibuprofen group were intraperitoneally injected with ibuprofen (30 mg/kg) once every one d 30 min before PTZ injection; rats in the control group were intraperitoneally injected with the same amount of normal saline every one d. Injection for 15 times was performed. After the last injection, the rats were observed for 10 min, and the latency, seizure level and complete ignition of the rats in each group were recorded. Electroencephalogram (EEG) was used to detect the abnormal brain discharge in rats. Four h after last injection, HE staining and Nissl staining were used to detect the proportion of damaged hippocampal neurons in each group. Immunohistochemical staining was used to detect the absorbance values of NLRP3 inflammasome, caspase-1 and interleukin (IL)-18 positive cells in the hippocampus of rats in each group; Western blotting was used to detect the protein expressions of NLRP3 inflammatome, caspase-1 and interleukin (IL)-18 in the hippocampus of each group. Results:(1) As compared with the PTZ group, rats in the PTZ+ibuprofen group had statistically lower incidence of complete ignition, significantly longer latency and significantly lower seizure level ( P<0.05). EEG showed spikes and high amplitude epileptic wave discharge in rats of the PTZ group; EEG showed low amplitude small spiny wave and slow spiny wave in rats of the PTZ+ibuprofen group. (2) As compared with the control group, the proportion of injured hippocampal neurons significantly increased in the PTZ group and PTZ+ibuprofen group ( P<0.05); and the proportion of injured hippocampal neurons in the PTZ+ibuprofen group signficantly decreased as compared with that in the PTZ group ( P<0.05). (3) As compared with those in the control group, the absorbance values of NLRP3 inflammatome, caspase-1 and IL-18 positive cells, and the protein expressions of NLRP3 inflammatome, caspase-1 and IL-18 in the hippocampus of the PTZ group and PTZ+ibuprofen group were all significantly increased ( P<0.05); as compared with the PTZ group, the the absorbance values of NLRP3 inflammatome, caspase-1 and IL-18 positive cells, and the protein expressions of NLRP3 inflammatome, caspase-1 and IL-18 in the hippocampus in the PTZ+ibuprofen group were all significantly decreased ( P<0.05). Conclusion:Ibuprofen can inhibit the expressions of NLRP3 inflammatome, caspase-1 and IL-18, reduce the intensity of seizures, and play a neuroprotective role.
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Objective To investigate the effect of ibuprofen on autophagy and astrocyte proliferation and their significances in rats with pentylenetetrazol (PTZ)-induced epilepsy. Methods Sixty male sprague-dawley rats were randomly divided into control group, PTZ group, 3-MA+PTZ group, ibuprofen+PTZ group and 3-MA+ibuprofen+PTZ group (n=12); activity of gamma-aminobutyric acid was blocked by PTZ to ignite epileptic rats in the latter 4 groups, and rats in the control group were intraperitoneally injected with normal saline once every one day; rats in the 3-MA+PTZ group and ibuprofen+PTZ group were given intraperitoneal injection of 3-MA (2.5 mg/kg) or ibuprofen (30 mg/kg) 30 min before PTZ injection;rats in the 3-MA+ibuprofen+PTZ group were given intraperitoneal injection of 3-MA (2.5 mg/kg)+ibuprofen (30 mg/kg) at the same time. The behavior and EEG features of rats were observed. Immunofluorescence staining and Western blotting were used to detect the expressions of microtubule-associated protein 1 light chain 3 (LC3) and glial fibrillary acidic protein (GFAP). Results (1) As compared with rats in the PTZ group and 3-MA+PTZ group, rats in the ibuprofen+PTZ group and 3-MA+ibuprofen+PTZ group had significantly decreased seizure grading and incidence of complete ignition, and significantly increased latency period (P<0.05). (2) The EEG waveform of the control group was normal; electroencephalogram of PTZ group and 3-MA+PTZ group showed sharp waves of high amplitude and spike waves; EEG wave peaks in the ibuprofen+PTZ group and 3-MA+ibuprofen+PTZ group decreased significantly, presenting frequent small spike waves and slow spike waves, among which ibuprofen+PTZ group showed more obvious changes. (3) The results of immunofluorescence staining and Western blotting showed that as compared with the PTZ group, 3-MA+PTZ group had statistically decreased LC3 expression and significantly increased GFAP expression (P<0.05); as compared with the PTZ group, ibuprofen+PTZ group had statistically increased LC3 expression and significantly decreased GFAP expression (P<0.05), however, 3-MA+ibuprofen+PTZ group had statistically decreased LC3 expression and significantly increased GFAP expression (P<0.05). Conclusion Ibuprofen can reduce astrocyte proliferation by promoting autophagy to affect seizures.
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Objective To explore the effect of quality control circle in decreasing the unqualified rate and improving quality of specimen in clinical laboratory during the pre-analytical phase.Methods The laboratory department of the First Affiliated Hospital of Kunming Medical University established quality control circle management model,since March 1 st to June 1 st 2016,following the steps of the theory and method of quality management.Used professional tools of quality control circle to solve existing problems before procedure.Then compared the disqualification rates before and after the implementation,and used chi-square test for statistical analysis.Results After activity of quality control circle,the unqualified rate of specimen in clinical laboratory declined from 0.565%to 0.220%.The difference was statistically significant (χ2=155.22,P<0.01), the goal achievement was 86.25%,the advance rate was 61.06%.And the ability of employees in using scientific means have improved.The ability of using quality control tools increased by 33.3%,the problem solving ability increased by 9.3%, the coordination and communication ability increased by 125%,the team cohesion increased by 56%and the enthusiasm increased by 20.7%. Conclusion With the application of quality control circle tools,the unqualified rate of specimen decreases and the ability of solving problems by applying of quality control circle tools.(Chin J Lab Med,2018,41:324-327)
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Almost 3% of newborns are diagnosed as small for gestational age(SGA)worldwide.Born SGA is one of the important causes of perinatal morbidity and mortality,and is also associated with metabolic diseases in adulthood.Low birth weight by itself is insufficient to characterize growth restriction,as it does not include information about the neonate’s body proportionality.Depending on the origin,timing and severity of in-sult,small for gestational age infants are classified into two types:proportionate or symmetric growth restriction (SGR)and disproportionate or asymmetric growth restriction(AGR).There may be differences in physical and neurological development of these two types.This study compares three classification indexes,and to find differ-ences in postnatal growth of these two types.
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Objective To investigate how to improve test quality by monitoring and analyzing 15 clinical laboratory quality indicators from the National Health and Family Planning Commission .Methods Data were collected from clinical laboratory department of the First Affiliated Hospital of Kunming Medical University between January 2011 and August 2015.15 quality indicators were analyzed retrospectively , including the error rate of specimen type , the coefficient variation unqualified rate of internal quality control test, the reporting rate of critical value , et al.Results The monitoring results of quality indicators basically satisfied the quality goals , except that the median of turn around time in pre-analytical phase was not established, routine internal quality control was not conducted in some laboratory tests in analytical phase and the reporting rate and reporting timely rate of critical value should be further improved in post -analytical phase .Conclusion Medical laboratory quality system can be continuously improved by means of setting up the quality goals of 15 quality indicators referring to sub-specialty and laboratory tests , as well as automated monitoring, statistics and analysis in LIS.
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Objective To evaluate the capability for detecting of tumor cells in ascites and pleural effusion by Sysmex XN‐2000 hematology analyzer .Methods Determination of 84 samples of ascites and pleural effusion specimens with Sysmex XN‐2000 hema‐tology analyzer humoral model to analyze the study parameters highly fluorescent cells absolute value (HF‐BF # ) and percentage (HF‐BF% ) of ascites and pleural effusions screening of tumor cells ,and related data for statistical analysis .Results In 84 cases of pleural effusion and ascites samples ,cytology found 14 cases of malignant cells ,cancer cells are not found in 70 cases .14 cases of malignant cases ,HF‐BF # average of 169 /μL(0-2 001) ,HF‐BF% with an average of 29 .7% (0% -261% );70 cases of benign cases ,HF‐BF # an average of 39 /μL(0-524) ,HF‐BF% with an average of 4 .2% (0% -27 .8% ) ,both groups were statistical differences in the mean significance(P<0 .05) .ROC curve analysis with high absolute value and percentage of fluorescent cells to tumor cells detection function ,HF‐BF # area under the curve(AUC) was 0 .493 ,the area(AUC)% HF‐BF under the curve was 0 .222 .Conclusion Sysmex XN‐2000 Pattern of humoral parameters of HF‐BF # and HF‐BF% ,although the tumor cells may pro‐vide some screening information ,but ineffective ,so we believe that in our daily work can′t be over‐reliance on the instrument HF‐BF# ,HF‐BF% and other parameters should be performed on every sample smear staining ,thus improving the detection rate of ma‐lignant cells .
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Objective To understand the distribution of overweight and obesity in 0-5 years old children who were born small for gestational age ( SGA) in Shanghai through a cross-sectional investigation. Methods All resident children aged 0-5 years were included, covering all 18 districts in Shanghai. Health-check cards were prepared for SGA children. The check-up included weight,length/height and head circumference. Body mass index ( BMI) was used to evaluate the overweight and obesity according to the standard of World Health Organization ( WHO) . Results This study investigated 728 602 children aged 0-5 years in Shanghai,and ultimately 23 871 of them were defined as SGA,a-mong whom 9 805(41. 4%) were boys and 14 066(58. 9%) were girls. The BMI of SGA children were higher than that of appropriate for gestational age( AGA) from 4 to 18 months,while for the rest of the time,they were basically the same. There was no difference in changing tendency of BMI between SGA children and those AGA children. The distri-bution of overweight and obesity according to the standard of WHO in SGA boys among different ages was 7. 7% to 20. 7%, and 15. 7% in average;the distribution of SGA girls among different ages was 5. 9% to 18. 3%,and 12. 9% in average. The proportion of overweight at the age of 4-18 months was significantly higher than that of other ages. There was no correlation between overweight at 0-2 years old and overweight at the age of 5(P>0. 05). Conclusions Com-pared with SGA girls,overweight and obesity in SGA boys were more serious. The age of 4-18 months were the period of high incidence of overweight. There was no correlation between overweight at 0-2 years old and overweight at the age of 5.
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Objective To analyze the infection of human parvovirus B19 among women of childbearing age in Xiangyang City, and to provide a reference for pregnant women's health care. Methods A total of 303 women of childbearing age in Xiangyang City from 2018 to 2019 were selected as the research subjects. B19 virus DNA in serum of the subjects was detected by nested PCR technology. The differences in the detection rate of B19 viral DNA among normal pregnancy, abnormal pregnancy, and infertility serum were statistically analyzed. The differences in the detection rate of B19 virus DNA among women of childbearing age at different ages were compared. Results The detection rate of B19 viral DNA in all 303 women of child-bearing age was 27.72%. The detection rate of B19 virus DNA in 26-35 year old women was higher than that in other age groups. The detection rate of B19 virus DNA in abnormal pregnancy group was significantly higher than that in normal pregnancy group (P <0.05). Conclusion The detection rate of B19 virus DNA in abnormal pregnancy and infertility group was significantly higher than that in normal pregnancy group, with the detection rate of B19 virus in 26-35 year old women of childbearing age being the highest among all age groups. It is necessary to strengthen the screening of B19 virus in pregnant women of childbearing age in this region to reduce its impact on fetal abortion.
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Objective: In order to evaluate the quality of Panax ginseng and its preparation, a simple and accurate HPLC method for determining the contents of 16 ginsenosides from P. ginseng was established. Methods: The chromatographic separation was achieved on a C18 column (150 mm × 4.6 mm, 5 μm) using a mobile phase made up of acetonitrie and water at a flow rate of 1.0 mL/min. The detection wavelength and column temperature were set as 203 nm and 35°C, respectively. Results: Sixteen ginsenosides (Rg1, Re, Rf, Rb1, Rg2, Rc, Rb2, Rb3, F1, Rd, F2, Rg3, protopanaxatriol, compound K, Rh2, and protopanaxadiol) were separated at baseline with good linearity (r ≥ 0.9990). The recovery rates were 95%-102% (RSD < 2%). Conclusion: The method is simple, fast, accurate, and could be applied to the quality control of P. ginseng and its preparation.
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A series of phthalazine ketone compounds were synthesized and the structures were confirmed by H NMR and HR-MS spectrum. All target compounds were obtained through 7 steps, including selective reduction, nitration, bromination, ring enlargement, reduction, Knoevenagel and acylated reaction. The compounds were evaluated for their immunosuppressive effects of T-cell proliferation and inhibitory activity of IMPDH type II in vitro, as well as their structure-activity relationship were assessed. Several compounds exhibited strong immunosuppressive properties, especially compounds 7f and 7h, with IC50 values of 0.093 micromol x L(-1) and 0.14 micromol x L(-1) respectively, which were superior to mycophenolic acid. The information obtained from the studies may be useful for further research on the immunosuppressive agents.
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Animals , Female , Mice , Cell Proliferation , IMP Dehydrogenase , Metabolism , Immunosuppressive Agents , Chemistry , Pharmacology , Inhibitory Concentration 50 , Mice, Inbred BALB C , Phthalazines , Chemistry , Pharmacology , Spleen , Cell Biology , Structure-Activity Relationship , T-LymphocytesABSTRACT
OBJECTIVE@#To investigate Homer protein expression after focal brain contusion and explore the relationship between expression and injury time.@*METHODS@#Focal brain contusion in rats was established and Homer protein expression in brain at different injury intervals after contusion was detected by immunohistochemistry and Western blotting.@*RESULTS@#A small amount of Homer positive expression cells were detected in control group, sham operated group and experimental group (0.5 h after contusion). The amount of Homer positive expression cells increased after 3 h and reached peak 12 h after contusion. The amount of positive cells continued to decrease 1 d after contusion and to the base level 7 d after contusion. Homer protein expression based on immunohistochemistry and Western blotting had statistical difference among adjacent groups.@*CONCLUSION@#Expression of Homer protein near the focal contusion area shows time dependence after brain contusion in rats.
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Animals , Male , Rats , Blotting, Western , Brain/pathology , Brain Injuries/pathology , Carrier Proteins/metabolism , Contusions/pathology , Disease Models, Animal , Forensic Pathology , Homer Scaffolding Proteins , Immunohistochemistry , Random Allocation , Rats, Wistar , Staining and Labeling , Time FactorsABSTRACT
<p><b>OBJECTIVE</b>To investigate the expression features of glypican-3 (GPC-3) and its diagnostic and differential values in hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>Rat hepatoma models were made and the dynamic expression features of GPC-3 protein and its gene were investigated by Western blotting and RT-PCR respectively. Liver specimens from 36 HCC patients were collected by self-control method and the expression and clinicopathological features of GPC-3 were analyzed by immunohistochemistry. Serum GPC-3 levels were quantitatively detected by ELISA and its efficiency for HCC diagnosis was evaluated in patients with liver diseases.</p><p><b>RESULTS</b>The incidence of GPC-3 was 0% in control, 83.3% in degeneration, 100% in precancerosis and 100% in canceration during dynamic formation of rat hepatoma, respectively. The positive GPC-3 was brown granule- like staining localized in membrane and cytoplasm in human HCC.</p><p><b>CONCLUSIONS</b>The GPC-3 positive rates were 80.6% in HCC, 41.7% in surrounding tissues and none in distal tissues (P < 0.01), respectively. No positive relationship presented between GPC-3 and differentiation grade or the number of tumor except of tumor size (Z = 2.941, P < 0.01). The incidence of serum GPC-3 was 52.8% in HCC patients except of one patient with cirrhosis. No significant differences were found between GPC-3 and sex, age, AFP, tumor number, Child classification or extrahepatic metastasis except of tumor size (χ² = 6.318, P < 0.05) and HBV infection (χ² = 23.362, P < 0.01). Combined detection of GPC-3 and AFP could rise up diagnosis of HCC. GPC-3 expression closely associated with HCC and might be useful for early diagnosis of HCC.</p>
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Adolescent , Adult , Aged , Aged, 80 and over , Animals , Female , Humans , Male , Middle Aged , Rats , Young Adult , Carcinoma, Hepatocellular , Diagnosis , Metabolism , Pathology , Diagnosis, Differential , Glypicans , Metabolism , Liver , Pathology , Liver Neoplasms , Diagnosis , Metabolism , Pathology , Rats, Sprague-DawleyABSTRACT
OBJECTIVE@#To investigate the expression of matrix metalloproteinase-3 after brain contusion and its applicability for estimating the age of brain contusion.@*METHODS@#Rats had been divided into three groups: control group, sham operation group and brain contusion group. The expression of matrix metalloproteinase-3 at different time was detected by immunohistochemistry and Western blot.@*RESULTS@#By the immunohistochemistry, no staining was observed in control and sham operation groups. The positive staining of MMP-3 appeared 6 hours after contusion, increased gradually in 24 hours and peaked 5 days after contusion, then started to decrease, 14 days after contusion still could be observed. By the Western blot analysis, no expression of MMP-3 was detected in control and sham groups. The positive staining of MMP-3 appeared 6 hours after contusion, increased gradually and maximized 5 days after contusion, then started to decrease, 14 days after contusion still could be found.@*CONCLUSION@#Time-order expression of MMP-3 could be used for estimating the age of brain contusion in forensic pathology.
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Animals , Male , Rats , Blotting, Western , Brain Injuries/enzymology , Forensic Pathology , Immunohistochemistry , Matrix Metalloproteinase 3/genetics , Random Allocation , Rats, Sprague-Dawley , Time FactorsABSTRACT
OBJECTIVE@#To establish a new animal model of grading skeletal muscle contusions that could be controllable and repetitive.@*METHODS@#The rats' gastrocnemius was injured by a new weight-dropping device designed. The force acting on gastrocnemius with a comparatively constant duration and inducing elastic deformation of the gastrocnemius was expressed with velocity (v) and deformation (DF). Instant velocity was changed to create gastrocnemius contusions. Pathological changes of gastrocnemius were graded by the gross and histological examinations of 39 rats.@*RESULTS@#At low level of impact (v: 2 m/s, DF: 5.5 mm), mild injuries were detected in epimysium and superficial layer of gastrocnemius. At moderate level of impact (v: 2.5 m/s, DF: 6.5 mm), the injuries were observed in epimysium and whole gastrocnemius. At high level of impact (v: 3 m/s, DF: 7.5 mm), severe injuries were seen deeper to soleus with more extensive skeletal muscle damage.@*CONCLUSION@#Grading of skeletal muscle blunt force contusion is created by parameter of velocity and muscle deformation. The model could be used for further research on skeletal muscle contusions.