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ObjectiveTo explore the effect of Tongxie Yaofang on the immune microenvironment of colorectal cancer in mice under chronic stress and the underlying mechanism. MethodA total of 40 male SPF BABL/C mice were randomized into normal group， stress group， Tongxie Yaofang group （13.65 g·kg-1）， and Tongxie Yaofang-stress group （13.65 g·kg-1）， with 10 in each group. Chronic restraint stress was induced in mice and administration （ig） of Tongxie Yaofang began after 7 days of stress. On the 14th day， forced swim and tail suspension tests were used to examine the behavioral changes of mice after stress and the subcutaneous colorectal tumor was implanted in each group of mice. The effect of this prescription on the body mass and tumor volume of mice was observed. After the last administration， mouse serum and tumor samples were collected. The content of T lymphocytes （CD3+， CD4+， CD8+， and CD4+/CD8+） in tumor was detected by immunohistochemistry and flow cytometry and levels of corticosterone （CORT） in peripheral blood， and interleukin （IL）-2， interferon-γ （IFN-γ）， IL-6， and IL-10 in the serum were determined by enzyme-linked immunosorbent assay （ELISA）. The protein expression of inhibitor of nuclear factor-κB（IκB） kinase α/β （IKKα/β）， nuclear factor-κB （NF-κB）α （IκBα）， NF-κB p65， and phosphorylated （p）-NF-κB p65 was measured by Western blot. ResultCompared with the normal group， the stress group had large tumor volume （P<0.05）， low content of CD3+， CD4+， and CD4+/CD8+ （P<0.05， P<0.01）， high content of CD8+， low content of T helper 1 （Th1）-secreted IFN-γ （P<0.05）， high content of T helper 2 （Th2）-secreted IL-10 （P<0.05） and CORT （P<0.05）， high protein expression of p-NF-κB p65， NF-κB p65， and IKKα/β （P<0.05）， and low protein expression of IκBα （P<0.05）. Compared with the normal group， the Tongxie Yaofang group showed slow tumor growth， high content of CD3+， CD4+， and CD4+/CD8+ （P<0.01）， low content of CD8+ （P<0.05）， high content of Th1-secreted IL-2 and IFN-γ （P<0.05）， low content of Th2-secreted IL-6 and IL-10 （P<0.05）， low content of CORT， low protein expression of p-NF-κB p65， NF-κB p65， and IKKα/β （P<0.05）， and high protein expression of IκBα （P<0.01）. Tongxie Yaofang-stress group demonstrated slower tumor growth， higher content of CD3+， CD4+， and CD4+/CD8+ （P<0.01）， smaller content of CD8+ （P<0.05）， higher content of IL-2 and IFN-γ （P<0.05）， lower content of IL-6， IL-10 （P<0.05）， and CORT （P<0.05）， lower protein expression of p-NF-κB p65， NF-κB p65， and IKKα/β （P<0.05，P<0.01）， and higher protein expression of IκBα （P<0.01） than the stress group. ConclusionTongxie Yaofang can delay the growth of colorectal cancer under chronic stress and alleviate the deterioration of the immune microenvironment， possibly by inhibiting NF-κB signaling pathway， regulating the function of T lymphocyte subsets， and thus suppressing the secretion of pro-inflammatory factors.
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Objective:To analyze the clinical characteristics and risk factors of impaired liver and renal function in hospitalized patients with gout.Methods:A total of 494 hospitalized patients with confirmed gout were selected and divided into four groups according to liver and renal function, control(Con), impaired liver function (ILF), impaired renal function (IRF), and both function impaired (ILRF) group. Multivariate logistic regression was used to analyze the risk factors related with impaired liver and renal function.Results:Compared to Con group, ILF group were younger with shorter gout duration, higher body mass index, waist circumference, homeostasis model assessment for insulin resistance (HOMA-IR), serum uric acid, low density lipoprotein-cholesterol (LDL-C), total cholesterol, triglycerides, C reactive protein, higher prevalence of dyslipidemia, obesity, fatty liver, and monosodium urate crystal (MSU) deposition (all P<0.05). IRF group were older and with higher serum uric acid, serum creatinine, C reactive protein, and hypertension, MSU deposition prevalence, with lower prevalence of fatty liver (all P<0.05). Compared to ILF group, IRF group were older, with longer gout duration, lower level of body mass index, waist circumference, HOMA-IR, LDL-C, total cholesterol, triglycerides, lower prevalence of obesity, fatty liver, and higher prevalence of hypertension and type 2 diabetes (all P<0.05). The univariate logistic regression analysis showed that age( OR=0.941, 95% CI 0.906-0.977, P<0.001), serum uric acid ( OR=1.002, 95% CI 1.000-1.005, P=0.043), HOMA-IR ( OR=1.147, 95% CI 1.024-1.285, P=0.018), and MSU deposition ( OR=1.959, 95% CI 1.154-3.326, P=0.013) were the independent risk factors of impaired liver function, while the independent risk factors of impaired renal function were age ( OR=1.104, 95% CI 1.048-1.162, P<0.001), serum uric acid ( OR=1.007, 95% CI 1.004-1.010, P<0.001), and MSU deposition ( OR=2.393, 95% CI 1.191-4.805, P=0.014). Conclusions:Serum uric acid and MSU deposition are the common independent risk factors for impaired liver and renal function in patients with gout. Younger patients with insulin resistance are susceptible to impaired liver function, older patients with hypertension and diabetes are susceptible to impaired renal function.
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Objective To analyze the clinical manifestation and CYP4V2 mutations of Bietti crystalline corneoretinal dystrophy( BCD) families. Methods Total of 234 patients (173 families) diagnosed as BCD were recruited in Peking University Third Hospital from 2010 to 2018. All of the subjects underwent comprehensive eye examinations to observe the clinical manifestations. Blood samples were collected and genomic DNA was extracted. The Sanger sequencing or high- throughput sequencing was applied for CYP4V2 gene mutation analysis. This study was approved by the Ethics Committee of Peking University Third Hospital (NO. 2012093). All patients and their family members signed informed consent. Results Some patients manifested the typical phenotype of BCD characterized by yellowish-white crystalline deposits throughout the fundus. However,some patients in advanced stage were easily misdiagnosed as other inherited retinal degeneration because the crystalline deposits diminished or even disappeared. Forty-nine probands in our cohort were misdiagnosed as other inherited retinal degeneration at first visit, with a misdiagnosis rate of 28. 3％. Genetic diagnosis results showed that 161 patients carried CYP4V2 mutation,and the positive rate was 93. 1％. Eight novel mutations were obtained. The three known mutations c. 802-8 _810del17bp, c. 1091-2 A>G and c. 992 A>C accounted for 73. 5％ of the mutations, which were hotspots in Chinese Han populations for BCD. Conclusions Patients with BCD have characteristic fundus manifestation, but are easily misdiagnosed in advanced stage. Molecular diagnosis is valuable in clinical diagnosis of the disease,thus contribute to the prevention and treatment of the disease. A single hybrid mutation is not enough to lead to BCD. No apparent genotype-phenotype correlation between the CYP4V2 gene and occurrence of BCD is identified.
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Objective@#To analyze the clinical manifestation and CYP4V2 mutations of Bietti crystalline corneoretinal dystrophy( BCD) families.@*Methods@#Total of 234 patients (173 families) diagnosed as BCD were recruited in Peking University Third Hospital from 2010 to 2018.All of the subjects underwent comprehensive eye examinations to observe the clinical manifestations.Blood samples were collected and genomic DNA was extracted.The Sanger sequencing or high- throughput sequencing was applied for CYP4V2 gene mutation analysis.This study was approved by the Ethics Committee of Peking University Third Hospital (NO.2012093). All patients and their family members signed informed consent.@*Results@#Some patients manifested the typical phenotype of BCD characterized by yellowish-white crystalline deposits throughout the fundus.However, some patients in advanced stage were easily misdiagnosed as other inherited retinal degeneration because the crystalline deposits diminished or even disappeared.Forty-nine probands in our cohort were misdiagnosed as other inherited retinal degeneration at first visit, with a misdiagnosis rate of 28.3%.Genetic diagnosis results showed that 161 patients carried CYP4V2 mutation, and the positive rate was 93.1%.Eight novel mutations were obtained.The three known mutations c. 802-8 _810del17bp, c.1091-2 A>G and c. 992 A>C accounted for 73.5% of the mutations, which were hotspots in Chinese Han populations for BCD.@*Conclusions@#Patients with BCD have characteristic fundus manifestation, but are easily misdiagnosed in advanced stage.Molecular diagnosis is valuable in clinical diagnosis of the disease, thus contribute to the prevention and treatment of the disease.A single hybrid mutation is not enough to lead to BCD.No apparent genotype- phenotype correlation between the CYP4V2 gene and occurrence of BCD is identified.
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BACKGROUND:Whether controling of post-injury inflammatory response combined with neural stem cel (NSC) transplantation can improve the curative efficacy for spinal cord injury stil remains unclear. OBJECTIVE:To investigate the repair of spinal cord tissue, myelin regeneration, axon regeneration, motor function recovery and the possible mechanism after early application of tumor necrosis factor α antagonist (Etanercept) combined with tyrosine kinase C (TrkC) gene-modified NSC transplantation. METHODS:TrkC-overexpressed NSCs (TrkC-NSCs) were constructed by lentiviral transfection technique. The rat models of spinal cord transection injury were prepared, and then subjected to Etanercept combined with TrkC-NSCs transplantation. The number of neurons and neuroregeneration after injury were measured by Nissl's staining, immunofluorescence and western blot. The rat motor function was detected by Basso Beattie Bresnahan score and evoked potential. The myelin regeneration was detected by electron microscopy and toluidine blue staining. RESULTS AND CONCLUSION:Compared with the other groups, the Etanercept combined with TrkC-NSCs transplantation group had more survived anterior horn motor neurons at 28 days after injury, more myelin-encapsulated axons, higher Basso Beattie Bresnahan score, greater amplitude of the evoked potentials, and relatively shorter latency (alP < 0.05). These findings indicate that early application of tumor necrosis factor-α antagonist combined with TrkC-NSCs transplantation after spinal cord injury in rats can effectively promote myelin regeneration, axon regeneration, and further promote motor function recovery.
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Objective To explore the neuroprotection and mechanisms of bone marrow mononuclear cells (BMMNCs),and evaluate whether ERK1/2 signaling pathway was involved in it.Methods384 healthy male SD rats,which were 6-8 week old,weighting 250-280 g,were selected.The middle cerebral artery occlusion (MCAO) model was established in SD rats using the suture method.The rats were randomly divided into sham operation group,model group,BMMNCs group and ERK1/2 inhibitor group,with 96 rats in each group.At the time of 24 h after the successful modeling,200 μl PBS solution was injected into the caudal vein of the rats in the model group,200 μl PBS solution containing 5×106 BMMNCs was injected into the rats in the BMMNCs group and the ERK1/2 inhibitor group.meanwhile,5 μl PD98059 was injected into the lateral ventricle of the brain of rats in the ERK1/2 inhibitor group.At the time points of 3 d,7 d and 14 d,the modified neurological severity scores (mNSS) was used to evaluate the neurological function,the volume of cerebral infarction was assessed by TTC staining,the pERK1/2,Bax,Bcl-2 and caspase-3 levels were detected by Western blot,and the effect of BMMNCs on activation of microglia was detected by immunofluorescence assay.Results(1)At each time point,the mNSS and the volume of cerebral infarction of the model group were significantly higher than those of the sham operation group (P0.05).(2)At each time point,the pERK1/2,Bcl-2,Bax and caspase-3 protein levels of the model group were significantly higher than those of the sham operation group (P0.05).(3) At each time point,microglia (Iba1 positive) in ischemic penumbra of the BMMNCs group was significantly more than those of the model group,and it was increased with the time extension (P0.05).ConclusionBMMNCs can reduce the apoptosis through ERK1/2 signaling pathway,thus improving the neurological function and reducing the infarct scope.
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Objective To investigate the clinical,imaging,pathological and molecular biological features of mitochondrial encephalomyopathy with lactic acidosis and stroke -like episodes(MELAS)in children.Methods The clinical,imaging,pathological and molecular biological features of 1 2 children with MELAS diagnosed through muscle biopsy or gene sequencing in the Fifth Affiliated Hospital of Zhengzhou University from January 201 1 to December 201 5 were retrospectively analyzed.Results (1 )Clinical features:the main manifestations included headache and vomiting in 1 1 cases,epileptic seizures in 9 cases,short stature in 8 cases,hairy in 7 cases,intolerance fatigue in 7 cases,cogni-tive decline in 7 cases,visual disturbance in 6 cases,hearing disturbance in 6 cases,and 5 cases had positive family history.In addition,7 cases had the serum lactic acid level increase in a rest for 1 0 min after exercise.(2)Imaging fea-tures:4 cases showed bilateral basal ganglia calcification symmetry in 8 patients who underwent head CT scan.The most frequently involved parts of the lesion were occipital in 1 0 cases,temporal in 9 cases and parietal lobe in 7 cases in stroke -like episodes.The lesions were lamellar necrosis.The abnormal areas by MRI showed low signal intensity on T1 weighted imaging,high signal intensity on T2 weighted imaging and fluid attenuated inversion recovery,high or equal signal intensity on diffusion weighted imaging,high or low signal intensity on apparent diffusion coefficient;the lactate peak significantly increased on magnetic resonance spectroscopy.The distribution was not in accordance with the control region of the cerebral vessels.Dynamic observation revealed that the lesions were reversible and migratory.(3)Myo-pathological features:muscle biopsy was performed in all children,and ragged -red fibers were found in 1 0 cases by im-proved Gomori staining,strongly succinate dehydrogenase -reactive were found in 9 cases,and the lipid droplets slight-ly increased in 8 cases by oil red O staining.Besides,the crystalline inclusion bodies in mitochondria were arranged in a parking lotpattern in 9 cases by electromicroscope.(4)Molecular biological characteristics:the mitochondrial gene mutations were analyzed in peripheral blood of 9 children and their parents,including 8 cases with A3243G muta-tion and 1 case with G13513A mutation.Five mothers had the same A3243G mutation site in 8 cases.Conclusions Children with MELAS have complex and varied clinical manifestations and certain characteristic of neuroimaging.More-over,muscle pathology and gene sequencing have important diagnostic value.Fully understanding the clinical,muscle pathology,imaging and molecular biological characteristics of children with MELAS can be helpful to the early diagnosis and treatment,also reduce misdiagnosis.
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ObjectiveTo investigate the mechanisms and the effects of magnesium Valproate on the expressions of the kinin B1 and B2 receptors in the hippocampus of the juvenile rats submitted to pilocarpine model of epilepsy.Methods 35 healthy Wistar juvenile rats were randomly divided into six groups,that is the model groups:Ⅰ group,Ⅱ group,Ⅲ group,and intraperitoneal injection of saline water control groups:Ⅰ a group,Ⅱ a group,Ⅲ a group,after succession of 15 rats to kindle to establish the model of epilepsy by pilocarpine.To collect hippocampus tissue after the rats were to put to death,and to compared the expression levels of kinin B1 and B2 receptor mRNA by RT-PCR and western blot in the hippocampus of rats.ResultsBy treated with magnesium valproate,kinin B1 receptor mRNA (0.38 ± 0.051 ) and protein expressions(0.58 ± 0.057 ) decreased and kinin B2 receptor mRNA (0.48 ±0.056 ) and protein expressions(0.48 ± 0.044 ) increased in Ⅰ group,compared with that (0.76 ±0.068,0.89 ± 0.034;0.28 ± 0.034,0.32 ± 0.039 ) of Ⅰ a group(P ＜ 0.05 ).Compared with control group,there were more significant upregulation of kinin B1 receptor mRNA and protein expressions (P＜0.05) in the Ⅰ and the Ⅱ groups and there were no alteration in Ⅲ group.The expressional levels of B2 receptor mRNA and protein were upregulated in the Ⅰ,Ⅱ and Ⅲ groups.ConclusionThe kinin B1 and B2 receptor may play a role in the onset and maintenance of epilepsy.The magnesium valproate increased the expressional levels of kinin B2 receptor,and decreased the expressional levels of kinin B1 receptor.
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In this study, real-time PCR and immunohistochemistry were used to detect coxsakie and adenovirus receptor (CAR) expression. Both localization and quantity were evaluated in the uteri obtained at days post coitus (dpc) 2.5, 4.5, 6.5, 8.5. Outcome of PCR was assessed by 2(-ΔΔCt) method. Image Pro-Plus 6.0 software was used for quantifying mean density of CAR expression in immunohistochemical sections. We found relatively weak CAR expression in the mouse uteri during implantation window. PCR and immunohistochemistry revealed highest CAR expression was detected on dpc 2.5 followed by down-regulation of CAR at dpc 4.5 and 6.5 (with significant difference). At dpc 8.5, CAR expression was increased slightly again. It is concluded that during implantation, the expression of CAR mRNA and protein is declined, resulting in the impairment of tight junction between cavity epithelium cells. After implantation window closure, CAR appears again to maintain epithelium stability. CAR might play an important role during embryo implantation procedure.
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Various collateral circulations were detected by 2D color Doppler flow imaging(2DCDFI)in the part of 70 patients of liver cirrhosis with portal hypertension,including reopened umbilical veins,dilated focal intrahepatic veins,dilating,hepatofugal centrifugal hepatic blood flow in the portalveins,perisplenic and retroperitoneai varices etc.Red and blue blood flows were detected in the collateral circulations by 2D-CDFI and appeared as continuous venous spectrum with low velocity blood volume could be calculated to evaluat the therapeulic effectiveness of portal-systemic shunt operation.
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Objective To assess the effect of arterial embolization in treating hyperthroidism by colour ultrasonography.Methods Forty two cases of hyperthyroidism were treated with thyroid arteries embolization. A few days before and 1, 3, 6 months after embolization, the echograms of thyroids were observed including the volumes of thyroids and the internal diameters of thyroid arteries were measured with colour ultrasonography respectively. The Vs, Vd, Vm, PI, RI were measured with the Doppler and the quantities of blood flow were calculated. The relationships of changes for all these parameters and T 3, T 4 and TSH were analysed. Results Before artery embolization all thyroids were enlarged with diffusely homogenous or heterogenous low echos and nodules in some patients accompanied by widenings of the thyroid arteries and their branches full of blood supply. The volume of thyroids decreased after artery embolization. 1 3 months later the echo of thyroids enhanced and got coarse with decrease of the thyroid vasculature and narrowing of vessel calibers except a few star or spot like blood streams were sometimes seen within the thyroids and no blood flow signals found in some cases. 3 6 months after embolization, the echo of thyroids decreased gradually or unevenly distribnted. All the parameters of blood flow before and after the treatment showed statistically significant differences( P