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ABSTRACT Objectives: Accurate preoperative prediction of adverse pathology is crucial for treatment planning of renal cell carcinoma (RCC). Previous studies have emphasized the potential of prostate-specific membrane antigen positron emission tomography / computed tomography (PSMA PET/CT) in differentiating between benign and malignant localized renal tumors. However, there is a scarcity of case reports elucidating the identification of aggressive pathological features using PET/CT. Our study was designed to prospectively compare the diagnostic value of enhanced CT, 68Ga-PSMA-11 and 18F-fluorodeoxyglucose (18F-FDG) PET/CT in clear-cell renal cell carcinoma (ccRCC) with necrosis or sarcomatoid or rhabdoid differentiation. Materials and Methods: A prospective case series of patients with a newly diagnosed renal mass who underwent enhanced CT, 68Ga-PSMA-11 and 18F-FDG PET/CT within 30 days prior to nephrectomy was included. Complete preoperative and postoperative clinicopathological data were recorded. Patients who received neoadjuvant targeted therapy, declined enhanced CT or PET/CT scanning, refused surgical treatment or had non-ccRCC pathological indications were excluded. Radiological parameters were compared within subgroups of pathological characteristics. Bonferroni corrections were used to adjust for multiple testing and statistical significance was set at a p-value less than 0.017. Results: Seventy-two patients were available for the final analysis. Enhanced CT demonstrated poor performance in identifying necrosis, sarcomatoid or rhabdoid differentiation and adverse pathology (all P > 0.05). The maximum standardized uptake value (SUVmax) of 68Ga-PSMA-11 PET/CT was more effective than 18F-FDG PET/CT in identifying tumor necrosis and adverse pathology, with an area under the curve (AUC) of 0.85 (cutoff value=25.26, p<0.001; Delong test z=2.709, p=0.007) for tumor necrosis and AUC of 0.90 (cutoff value=25.26, p<0.001; Delong test z=3.433, p<0.001) for adverse pathology. However, no significant statistical difference was found between 68Ga-PSMA-11 and 18F-FDG PET/CT in predicting sarcomatoid or rhabdoid feature (AUC of 0.91 vs.0.75, Delong test z=1.998, p=0.046). Subgroup analyses based on age, sex, tumor location, maximal diameter, stage and WHO/ISUP grade demonstrated that 68Ga-PSMA-11 PET/CT SUVmax had a significant predictive value for adverse pathology. Enhanced CT value and SUVmax demonstrated strong reliability [intraclass correlation coefficient (ICC) > 0.80], indicating a robust correlation. Conclusions: 68Ga-PSMA-11 PET/CT demonstrates distinct advantages in identifying aggressive pathological features of primary ccRCC when compared to enhanced CT and 18F-FDG PET/CT. Further research and assessment are warranted to fully establish the clinical utility of 68Ga-PSMA-11 PET/CT in ccRCC.
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The aim of this study was to investigate the role and mechanism of terpinen-4-ol (T4O) on high glucose (HG) -induced calcification in vascular smooth muscle cell (VSMC). To investigate the role of T4O on HG-induced calcium deposition, osteogenic phenotypic transformation and mitochondrial dynamics in VSMC, Mdivi-1, a mitochondrial dynamin-related protein 1 (Drp-1) inhibitor, was used to analyze the correlation between mitochondrial dynamics and VSMC calcification and the role of T4O. Alizarin red S staining was used to observe calcium salt deposition and flow cytometry to detect intracellular Ca2+ content; Western blot and immunofluorescence were used to detect the expression of phenotypic switching-related markers α-smooth muscle actin (α-SMA), bone morphogenetic protein 2 (BMP2) and Runt related transcription factor 2 (Runx2), and mitochondrial dynamics-related markers mitofusin 1 (MFN1), mitofusin 2 (MFN2) and Drp-1. The results showed that low and high doses of T4O could inhibit HG-induced down-regulation of α-SMA, MFN1 and MFN2 expression levels, and up-regulation of BMP2, Runx2 and Drp-1 expression levels, reduce intracellular Ca2+ content and calcium salt deposition, and effectively inhibit HG-induced VSMC calcification and mitochondrial dynamics disorders. The T4O group, Mdivi-1 group and T4O+Mdivi-1 group were able to up-regulate the expression levels of HG-induced α-SMA, MFN1 and MFN2, down-regulate the protein expression levels of BMP2, Runx2 and Drp-1, and inhibit calcium salt deposition, and there was no significant difference between the above indexes in the T4O and T4O+Mdivi-1 groups. The above findings suggest that T4O can inhibit the expression level of Drp-1, regulate the disturbance of mitochondrial dynamics, and suppress HG-induced VSMC calcification.
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Background Osteoclast stimulatory transmembrane protein (OC-STAMP) is involved in silicosis fibrosis induced by silicon oxide (SiO2) exposure. Its role in silicosis fibrosis by inducing ferroptosis of alveolar type II epithelial cells and its related mechanism remain unclear. Objective To explore the effect and possible mechanism of OC-STAMP on ferroptosis of alveolar type II epithelial cells and silicosis fibrosis in rats under SiO2 exposure. Methods Twenty male Wistar rats of SPF grade were randomly divided into two groups: control (Sham) group and SiO2 group, 15 rats in each group. Rats in the SiO2 group were given 1 mL of 50 mg·L−1 SiO2 suspension at one time through the non-exposed intratracheal instillation method to establish an animal model of silicosis, and rats in the Sham group were give 1 mL of 0.9% sodium chloride solution in the same way. Rats were sacrificed after 8 weeks. Samples of lung tissue were fixed in glutaraldehyde or paraformaldehyde for observing ultrastructure of mitochondria by transmission electron microscopy; HE, Masson, VG, and Prussian blue were used to observe changes in lung tissue structure and iron deposition. The expression level of OC-STAMP and the degree of lung fibrosis were evaluated by immunohistochemistry and immunofluorescence. The expression level of OC-STAMP in rat lung tissue was detected and the transfection effect of OC-STAMP was verified by real-time fluorescence quantitative polymerase chain reaction (RT-PCR). Overexpression (OCS group) and inhibition expression (SI-OC group) models were constructed by OC-STAMP plasmid and OC-STAMP small interfering RNA (siRNA) transfection to cultured MLE-12 cells, respectively. The relative expression levels of glutathione peroxidase 4 (GPX4), solute carrier family 7 member 11 (SLC7A11), and other proteins in lung tissue and MLE-12 were detected by Western blotting. Results The results of HE, Masson, and VG staining showed that the silicosis modeling was successful after 8 weeks of SiO2 exposure. The immunofluorescence results showed that OC-STAMP and ATP binding cassette subfamily A member 3 (ABCA3) co-localized in alveolar type II epithelium. The immunohistochemical results showed that the levels of OC-STAMP and collagen I in the SiO2 group were significantly higher than those in the Sham group (P<0.01). The RT-PCR results showed that the OC-STAMP mRNA in the lung tissue of the SiO2 group was significantly higher than that of the Sham group (P<0.01). The Prussian blue staining in the lung tissue of the SiO2 group showed positive brownish-yellow particles. Compared with the Sham group which showed normal mitochondrial structure, the mitochondrial structure was generally swollen and the mitochondrial cristae dissolved and disappeared in the SiO2 group by transmission electron microscope observation. The Western blotting results showed that the expression levels of SLC7A11 and GPX4 both decreased in the lung tissue of the SiO2 group (P<0.05, P<0.01), and the expression level of Vimentin increased (P<0.01). In the transfected MLE-12 cells, compared with the Sham group, the expression levels of SLC7A11 and GPX4 in the OCS group were significantly reduced (P<0.05, P<0.01). Conclusion OC-STAMP may affect the expression of proteins related to ferroptosis, and promote lung fibrosis induced by SiO2 exposure.
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Objective:To investigate clinical and laboratory characteristics of secondary hemophagocytic lymphohistiocytosis (sHLH) associated with secondary cutaneous T-cell lymphoma (CTCL) .Methods:CTCL patients with clinically suspected sHLH were collected from Department of Hematology, Wuhan No.1 Hospital from January 2016 to October 2021, and were evaluated according to the HLH-2004 diagnostic criteria and HScore.Results:Seven CTCL patients were confirmedly diagnosed with sHLH, including 2 with primary cutaneous γδT-cell lymphoma (PC-GDTCL) , 3 with cutaneous extranodal natural killer/T-cell lymphoma (C-ENKTCL) , and 2 with primary cutaneous anaplastic large cell lymphoma (PC-ALCL) . All the 7 patients received chemotherapy, but 6 died finally, and the median overall survival duration was 26.5 days (range: 14 - 60 days) after the confirmed diagnosis of CTCL complicated by sHLH. HLH-related gene mutations, which were located in the PRF1 and LYST genes, were identified in 2 patients; lymphoma-related gene mutations were identified in the KRAS and KMT2D genes in 1 PC-GDTCL patient,and in the JAK3 and SAMHD1 genes in another PC-GDTCL patient.Conclusions:CTCL complicated by sHLH usually progresses rapidly, so early diagnosis and treatment are needed. Bone marrow biopsy and mutation screening of lymphoma- and HLH-related genes at initial diagnosis and during disease progression may facilitate early diagnosis.
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Silicosis is a common occupational disease caused by long-term inhalation of large amounts of free SiO2 dust and deposition in lung tissues, characterized by the formation of silicon nodules and diffuse fibrosis of lung tissues. Silicosis is one of the most common and serious occupational diseases in China, and its treatment imposes a huge economic burden on individuals and the country. The formation mechanism of silicosis is very complex, and no early screening indicators, effective drugs, and treatment methods are available yet. The current diagnosis of silicosis is based on occupational history and chest radiography findings, and it is irreversible once pulmonary fibrosis develops. Moreover, as silicosis is a continuously progressive disease, even if silicosis patients stop exposure to free SiO2 dust, their pulmonary fibrosis will continue to develop and deteriorate. Programmed cell death (autophagy, apoptosis, ferroptosis, etc.) is a key factor involved in the development of silicosis. This article summarized the important roles of programmed cell death, including autophagy, apoptosis, and ferroptosis, in silicotic fibrosis, and concluded that regulating different programmed cell death and related signaling pathways through effective means may delay the process of silicosis fibrosis, providing new ideas and clues for exploring potential mechanisms of silicosis formation and formulating prevention and treatment strategies.
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Objective: To investigate the clinical and biological characteristics of familial platelet disorder (FPD) with germline Runt-related transcription factor (RUNX) 1 mutations. Methods: Patients diagnosed with myelodysplastic syndrome (MDS) or acute myeloid leukemia (AML) with RUNX1 mutations from February 2016 to December 2021 in Wuhan No.1 Hospital underwent pedigree analysis and were screened for gene mutations (somatic and germline). Patients diagnosed with FPD with germline RUNX1 mutations were enrolled and evaluated in terms of clinical characteristics and biological evolution. Bioinformatics analysis was used to assess the pathogenicity of mutations and to analyze the effect of mutated genes on the function of the corresponding protein. Results: Germline RUNX1 mutations were detected in three out of 34 patients suffering from MDS/AML who had RUNX1 mutations. A pedigree of FPD with RUNX1 (RUNX1-FPD) c.562A>C and RUNX1 c.1415T>C mutations was diagnosed, and the mutations were of patrilineal origin. Bioinformatics analysis indicated that the locus at positions 188 and 472 in the AML-1G type of RUNX1 was highly conserved across different species, and that variations might influence functions of the proteins. The mutations were evaluated to be highly pathogenic. Of the nine cases with germline RUNX1 mutations: two patients died due AML progression; one case with AML survived without leukemia after transplantation of hemopoietic stem cells; four patients showed mild-to-moderate thrombocytopenia; two cases had no thrombocytopenia. During the disease course of the proband and her son, mutations in RUNX1, NRAS and/or CEBPA and KIT appeared in succession, and expression of cluster of differentiation-7 on tumor cells was enhanced gradually. None of the gene mutations correlated with the tumor were detected in the four cases not suffering from MDS/AML, and they survived until the end of follow-up. Conclusions: RUNX1-FPD was rare. The mutations c.562A>C and c.1415T>C of RUNX1 could be the disease-causing genes for the family with RUNX1-FPD, and these mutations could promote malignant transformation. Biological monitoring should be carried out regularly to aid early intervention for family members with RUNX1-FPD.
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Humanos , Femenino , Mutación de Línea Germinal , Subunidad alfa 2 del Factor de Unión al Sitio Principal/genética , Linaje , Trastornos de las Plaquetas Sanguíneas/complicaciones , Leucemia Mieloide Aguda/genéticaRESUMEN
This study aims to investigate the efficacy and possible mechanism of Liuwei Dihuang Pills in the treatment of diminished ovarian reserve(DOR) by using proteomic techniques. Firstly, cyclophosphamide(60 mg·kg~(-1)) combined with busulfan(6 mg·kg~(-1)) was injected intraperitoneally to establish the mouse model of DOR. After drug injection, the mice were continuously observed and the success of modeling was evaluated by the disturbance of the estrous cycle. After successful modeling, the mice were administrated with the suspension of Liuwei Dihuang Pills by gavage for 28 days. At the end of the gavage, four female mice were selected and caged together with males at a ratio of 2∶1 for the determination of the pregnancy rate. Blood and ovary samples were collected from the remaining mice on the next day after the end of gavage. Hematoxylin-eosin(HE) staining and transmission electron microscopy(TEM) were then employed to observe the morphological and ultrastructural changes in the ovaries. The serum levels of hormones and oxidation indicators were measured by enzyme-linked immunosorbent assay. Quantitative proteomics techniques were used to compare the ovarian protein expression before and after modeling and before and after the intervention with Liuwei Dihuang Pills. The results showed that Liuwei Dihuang Pills regulated the estrous cycle of DOR mice, elevated the serum levels of hormones and anti-oxidation indicators, promoted follicle development, protected the mitochondrial morphology of ovarian granulosa cells, and increased the litter size and survival of DOR mice. Furthermore, Liuwei Dihuang Pills negatively regulated the expression of 12 differentially expressed proteins associated with DOR, which were mainly involved in lipid catabolism, inflammatory response, immune regulation, and coenzyme biosynthesis. These differentially expressed proteins were significantly enriched in sphingolipid metabolism, arachidonic acid metabolism, ribosomes, ferroptosis, and cGMP-PKG signaling pathway. In summary, the occurrence of DOR and the treatment of DOR with Liuwei Dihuang Pills are associated with multiple biological pathways, mainly including oxidative stress response, inflammatory response, and immune regulation. "Mitochondria-oxidative stress-apoptosis" is the key to the treatment of DOR by Liuwei Dihuang Pills. YY1 and CYP4F3 may be the key upstream targets that trigger mitochondrial dysfunction and ROS accumulation, and the metabolism of arachidonic acid is the main signaling pathway of drug action.
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Femenino , Masculino , Embarazo , Animales , Ratones , Ácido Araquidónico , Reserva Ovárica , Proteómica , Ovario , Metabolismo de los LípidosRESUMEN
Objective:To study the effect of the reform of pathology teaching mode.Methods:The Batch 2017 Class 1 and Class 3 students of clinical medicine who had pathology courses in the second semester of the 2019-2020 school year were selected in the controlled study, and they were divided into the study group (56 students from Class 1) and the control group (57 students from Class 3). For the pathology course, the control group used conventional teaching, and the study group used online-offline mixed teaching combined with problem-based learning (PBL). Both groups were taught for 1 semester. The theoretical assessment scores, practical assessment scores and excellent rate of the two groups after teaching were compared, the abilities of autonomous learning, problem solving, teamwork, and multidisciplinary thinking were compared between the two groups before and after teaching, and the satisfaction with the teaching mode was compared between the two groups. SPSS 23.0 software was used for t test and chi-square test. Results:The theoretical assessment scores [(93.86±5.42) vs. (86.74±5.33)] and practical assessment scores [(92.91±5.37) vs. (84.86±5.26)] of the study group were significantly higher than those of the control group ( P<0.05); the differences in grades distribution were statistically significant, and the excellent and good rate of the study group was higher than that of the control group ( P<0.05). The scores of autonomous learning, problem solving, teamwork and multidisciplinary thinking in the two groups were higher than those before the teaching, and the scores of the study group were higher than those of the control group ( P<0.05); after the teaching, the study group had higher satisfaction scores than the control group in enhancing clinical thinking ability, deepening the perception of life value and other aspects ( P<0.05). Conclusion:The combination of online and offline teaching combined with PBL can not only improve the assessment performance and excellent rate of medical students majoring in clinical medicine, but also enhance the ability of medical students to study independently, solve problems, teamwork and multidisciplinary thinking, and improve their satisfaction with the teaching mode.
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OBJECTIVE@#To establish an immune gene prognostic model of acute myeloid leukemia (AML) and explore its correlation with immune cells in bone marrow microenvironment.@*METHODS@#Gene expression profile and clinical data of TCGA-AML were downloaded from TCGA database. Immune genes were screened by LASSO analysis to construct prognosis prediction model, and prediction accuracy of the model was quantified by receiver operating characteristic curve and area under the curve. Survival analysis was performed by Log-rank test. Enriched pathways in the different immune risk subtypes were evaluated from train cohort. The relationship between immune prediction model and bone marrow immune microenvironment was verified by flow cytometry in the real world.@*RESULTS@#Patients with low-risk score of immune gene model had better prognosis than those with high-risk score. Multivariate analysis showed that the immune gene risk model was an independent prognostic factor. The risk ratio for AML patients in the training concentration was HR=24.594 (95%CI: 6.180-97.878), and the AUC for 1-year, 3-year, and 5-year overall survival rate was 0.811, 0.815, and 0.837, respectively. In addition, enrichment analysis of differential gene sets indicated activation of immune-related pathways such as cytokines and chemokines as well as autoimmune disease-related pathways. At the same time, real world data showed that patients with high immune risk had lower numbers of CD8+T cells and B lymphocytes compared with low immune risk patients.@*CONCLUSION@#We constructed a stable prognostic model for AML, which can not only predict the prognosis of AML, but also reveal the dysregulation of immune microenvironment.
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Humanos , Leucemia Mieloide Aguda/genética , Pronóstico , Curva ROC , Factores de Riesgo , Transcriptoma , Microambiente Tumoral/genéticaRESUMEN
Objective:To investigate the effect of hydroxysafflor yellow A(HSYA) preconditioning group on apoptosis induced by cold hypoxia/reoxygenation (cold H/R) injury in human renal tubular epithelial cells (HK2 cells).Methods:After digestion and passage, HK2 cell lines were divided into Sham group (control group), cold hypoxia and reoxygenation group (cold H/R group, cells cold hypoxia for 4 h, reoxygenation for 4 h), and HSYA preconditioning group (each HSYA subgroup was given different doses of HSYA 0.5 h before hypoxia, and the other operations were the same as the cold H/R group). The cell survival rate was measured by CCK-8 method.The expression of Bcl-2, Bax and Caspase-3 proteins in HK-2 cells were detected by immunocytochemistry and Western blotting.Results:(1) Compared with cold H/R group, different doses of HSYA could improve cell survival rate in different degrees, but only HSYA25 μmol/L group had the most significant effect (74.000±5.500 vs.59.000±3.800, P<0.05). (2) Immunocytochemistry semi-quantitative score: Compared with cold H/R group, the expression of Bax and Caspase-3 in HK2 cells of HSYA25 μmol/L group was significantly decreased(0(0, 1) vs. 8(6, 8), Z=2.041, P<0.05 and (3.400±0.548) vs.(7.800±1.095), t=11.000, P<0.01). The expression of Bcl-2 protein was increased significantly ((6.800±1.095) vs.(1.400±0.548), t=10.590, P<0.01). The ratio of Bcl-2/Bax increased significantly.(3)Western blot was used to detect protein: Compared with the cold H/R group, the protein levels of Bax, Cleaved-Caspase-3 and Pro-caspase-3 of HK2 cells in the HSYA25 μmol/L group were significantly decreased ((0.707±0.012) vs.(0.968±0.117), (0.480±0.009)vs.(0.735±0.005), (0.992±0.008)vs.(1.197±0.005), all P<0.01). The expression of Bcl-2 protein was significantly increased, and the ratio of Bcl-2/Bax was significantly increased ((0.410±0.009) vs.(0.273±0.008), (0.582±0.016) vs (0.282±0.080), all P<0.01). The experimental results were consistent with the immunocytochemistry. Conclusion:HSYA can effectively reduce the damage of HK2 cells after cold hypoxia and reoxygenation.
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Objective:To explore the effect of modified Chaiqin Wendantang on blood glucose level and gastrointestinal dysfunction in patients with diabetic gastroparesis (DGP) of weak spleen and stomach. Method:A total of 138 patients with DGP of weak spleen and stomach in Hainan Provincial Fourth People's Hospital from February 2017 to March 2019 were enrolled, and divided into two groups according to the random number table methods. Both groups received the routine treatment. In addition to this, study group received Chaiqin Wendantang, while control group received domperidone tablets. Traditional Chinese medicine (TCM) syndrome scores, blood glucose, gastrointestinal function, hemorheology index, gastric emptying function and gastric electrical activity, Pittsburgh Sleep Quality Index (PSQI) scale and clinical efficacy were compared. Result:After treatment, TCM symptom scores and total scores decreased (P<0.05), levels of fasting blood glucose (FBG), 2 hours postprandial blood glucose (2 h PBG), and glycated hemoglobin (HbA1c) decreased (P<0.05), serum motilin (MOT) and gastrin (GAS) levels increased (P<0.05), cholecystokinin (CCK) levels decreased (P<0.05), gastric emptying time shortened, frequency, amplitude and rhythm increased (P<0.05), PSQI score decreased (P<0.05), and whole blood viscosity (WBV) and fibrinogen (FIB) levels decreased (P<0.05), all of those changes were more obvious in study group than control group (P<0.05). The total effective rate in study group was higher than that in control group (P<0.05). During the treatment period, there were no obvious adverse reactions in study group, while there were 2 cases of transient dizziness and headache in control group, which were relieved after several seconds. The recurrence rate in study group was lower than that in control group (P<0.05). Conclusion:Modified Chaiqin Wendantang can effectively ameliorate the symptoms of gastric retention, improve sleep quality, control blood glucose levels, and improve hemodynamics for DGP of weak spleen and stomach patients. Besides, it can improve the gastrointestinal function by reducing serum CCK levels, so as to stimulate the secretion of MOT and GAS, increase gastric motility, shorten gastric emptying time, and promote the recovery of gastric electrical activity. With a high safety and low recurrence rate, it has clinical application value.
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Objective@#To investigate the influence of the degrees of intravesical prostatic protrusion (IPP) on the recovery of urinary continence after radical prostatectomy.@*METHODS@#We retrospectively analyzed the clinical data on 212 patients diagnosed with prostate cancer by biopsy and treated by laparoscopic radical prostatectomy by the same surgeon. Based on the degrees of IPP measured by MRI, we divided the patients into an IPP ≤ 10 mm group (n = 146) and an IPP > 10 mm group (n = 66) and determined the factors influencing the recovery of urinary continence by univariate and multivariate logistic regression analyses.@*RESULTS@#At 1, 3, 6 and 12 months after surgery, the urinary continence rates of the patients were 32.5%, 50.5%, 82.1% and 91%, respectively. Univariate analysis indicated that the factors influencing the recovery of urinary continence included IPP, body mass index (BMI), bladder neck preservation (BNP), neurovascular bundle preservation (NVBP) and clinical tumor (T) stage at 3 months (P 10 mm (P 10 mm and BMI ≥ 25 kg/m2 are independent factors influencing the long-term recovery of urinary continence after radical prostatectomy.
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@#Diabetic retinopathy(DR), one of the most common complications of diabetes mellitus, is the leading cause of blindness in working-age population. DR, previously regarded as a microvascular disease, is also considered as neuronopathy and low-to-moderate inflammation in retina with research progression. Microglias, the resident macrophage in the inner retina, are responsible for surveillance of the microenvironment in retina. Under abnormal conditions, microglias are activated and interact with different types of cells in retina. In DR, microglias become activated, as evidenced by the activation of the key molecules or signal transduction pathways, such as the nuclear factor kappa-light-chain-enhancer of activated B cells(NF-κB)and extracellular signal-regulated kinase(ERK)signaling pathways, which lead to the increased production of pro-inflammatory factors, chemokines,<i> etc.</i> At the same time, the proliferation and migration of activated microglia are enhanced, and microglias migrate to the outer retina. The over-activation of microglias causes neuronal cell apoptosis and blood-retinal barrier breakdown, resulting in vision loss.
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OBJECTIVE: To observe the effect of manual acupuncture stimulation of different layers (skin, muscle, peritoneum, sub-peritoneum) of "Tianshu" (ST 25) region on proximal colonic pressure in normal rats. METHODS: Forty-eight male SD rats were divided into 6 groups: all layer-needling, brushing, cutaneous needling, muscular needling, peritoneum-needling and sub-peritoneum-needling groups (n＝8 in each group). Manual needling or brushing was applied to "Tianshu" (ST 25) region. The colonic internal pressure was measured by using an amplifier and a pressure transducer-connected balloon which was implanted into the colonic cavity about 6 cm from the ileocecal valve. For rats of the all-layer needling group, an acupuncture needle was inserted into ST 25 about 1 cm deep and rotated for a while, for rats of the brushing group, a Chinese calligraphy brush pen was used to brush the skin hair for 1 min. For rats of the rest 4 groups, an acupuncture needle was inserted into the skin, muscle layer after cutting open the skin (about 0.1 cm), the peritoneum layer after cutting open the skin and muscle layers, and the sub-peritoneum layer after cutting open the skin, muscle and peritoneum layers, respectively, and rotated using the uniform reinforcing-reducing technique for about 1 min at a frequency of 120 twirlings per minute every time. RESULTS: During manual needling stimulation of the full layers, cutaneous layer, muscle layer, peritoneum layer and the sub-peritoneum layer of bilateral "Tianshu" (ST 25), the internal pressure of proximal colon was significantly decreased relevant to pre-stimulation in each group (P0.05). During hair brushing of ST 25 region, the colonic pressure was observably increased relevant to pre-needling stimulation (P<0.05). One min after the acupuncture stimulation, the decreased pressures maintained in needling the all-layer on the left side, needling the skin on the right side, needling the peritoneum layer on both sides, and needling the sub-peritoneum layer on both sides relevant to the brushing group of the same side (P<0.05). CONCLUSION: Manual acupuncture stimulation of each layer tissue of ST 25 on both sides may lower internal pressure of proximal colon in normal rats, suggesting their involvement of acupuncture effect in relaxing proximal colonic contraction.
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An analytical method was developed for determination of ginkgolic acids in Yinxing Tongzhi Dropping Pills by ultra high performance liquid chromatography-triple quadrupole mass spectrometry. The samples were purified by mix-mode anion exchange and reversed-phase SPE. A chromatographic column, Waters Cortecs T3 (50 mm×2.1 mm, 2.7 μm), was used with acetonitrile-methanol-1% acetic acid (44:44:12) as the mobile phase. The ginkgolic acids were detected by electrospray ionization mass spectrometry in negative mode with multiple reaction monitoring (MRM) mode. Ginkgolic acid C13:0, C15:1 and C17:1 possessed good linear correlation in the mass concentration range from 0.2 to 200 μg·L-1, 2 to 200 μg·L-1, 4 to 200 μg·L-1, respectively, with the correlation coefficients more than 0.999. The mean recoveries at spiked levels of 50, 250 and 600 μg·kg-1 were in the range of 70.8%-95.1%, and the RSDs were 0.7%-8.6%. The limits of quantification were 1, 10, 20 μg·kg-1, respectively. The method could be applied to the analysis of ginkgolic acids in complex matrix samples.
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Objective To observe the effect of HIF-1a/iNOS signaling pathway on myocardial ischemia-reperfusion injury in rat heart transplantation and the protective mechanism of N-acetylcysteine (NAC) on donor heart after cardiac transplantation in rats.Methods Eighty healthy male Lewis rats were randomly divided into 3 groups,the control group (0.3 ml saline was infused via inferior vena cava 30 min before donor harvest or implantation),NAC donor pretreatment group [NAC (30 mg/kg.w) was injected into the vena cava of donor rat 30 min defore donor harvest],and the NAC receptor pretreatment group (NAC 300 mg/kg.w was injected into the vena cava of the recipient rats 30 min before transplantation.The 30 min was injected into the vena cava of the recipient rats).A transplant model was established and the graft was obtained after 24 h transplantation.The expression of iNOS,HIF-1a and mRNA in cardiac muscle tissue was detected by immunohistochemistry and Real time-PCR.Results HIF-1a protein expression in graft myocardial tissue was significantly lower in NAC donor pretreatment and recipient pretreatment group compared with control group (P ＜0.05),the differences were statistically significant (2.72±0.17 vs.2.24±0.23 vs.3.14.±0.16,F=56.26,P =0.000).The iNOS protein expression in NAC donor pretreatment group,and NAC recipient pretreatment group were lower than that in the control group (1.52±0.18 vs.1.61±0.19 vs.3.30±0.18,F=232.345,P =0.000),the differences were statistically significant (P ＜ 0.05).24 h after transplantation,the differences in graft myocardial tissue HIF-1a and iNOS mRNA among the three groups were statistically significant (F=7.467,16.490,P=0.003,0.000).The expression of iNOS mRNA in the NAC receptor pretreatment group was significantly lower than that in the control group (P＜0.05).Conclusion HIF-1a/iNOS signaling pathway can regulate ischemia reperfusion injury in rat heart transplantation,and the protective effect of NAC on donor heart maybe mediated via this pathway.
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The present study was aimed to investigate the electrophysiological characteristics of hippocampal postnatal early development mediated by α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors in rats. Forty-eight Wistar rats were divided into postnatal 0.5-, 1-, 2- and 3-month groups (n = 12). Spontaneous excitatory postsynaptic currents (sEPSCs) and field excitatory postsynaptic potentials (fEPSPs) mediated by AMPA receptors were recorded to evaluate the changes in the intrinsic membrane properties of hippocampal CA1 pyramidal neurons by using patch-clamp and MED64 planar microelectrode array technique respectively. The results showed that, during the period of postnatal 0.5-3 months, some of the intrinsic membrane properties of hippocampal CA1 pyramidal neurons, such as the membrane capacitance (Cm) and the resting membrane potential (RMP), showed no significant changes, while the membrane input resistance (Rin) and the time constant (τ) of the cells were decreased significantly. The amplitude, frequency and kinetics (both rise and decay times) of sEPSCs were significantly increased during the period of postnatal 0.5-1 month, but they were all decreased during the period of postnatal 1-3 months. In addition, the range of evoked fEPSPs in hippocamal CA1 region was significantly expanded, but the fEPSP amplitudes were decreased significantly during the period of postnatal 0.5-3 months. Furthermore, the evoked fEPSPs could be significantly inhibited by extracellular application of the AMPA receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX). These results suggest that AMPA receptor may act as a major type of excitatory receptor to regulate synaptic transmission and connections during the early stage of hippocampal postnatal development, which promotes the development and functional maturation of hippocampus in rats.
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Coding rules for Chinese medicines and their codes (GB/T 31774-2015) was issued by General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China (AQSIQ) and Standardization Administration of the People's Republic of China (SAC) in 2015. Coding system for Chinese medicines (ISO 18668-1, 2, 3 and 4) series were issued one after another by International Organization for Standardization in 2016 and 2017. In this paper, the comparative study on the GB/T 31774 and ISO 18668-1, 2, 3 to 4 would be conducted to expound the similarities and differences among them. This essay aims at promoting the application of national and international standards of coding system in production and operation enterprises and the medical institutions of traditional Chinese medicine (TCM), reducing their repetitive investment to meet the Chinese medicine import and export trade requirement in future. Moreover, it provides the cornerstone and support for TCM standardization, and makes Chinese medicines standard gain dominance in field of international TCM standards, occupying the high ground of international market in the traditional medicine field of the world. It may promote the "Internet + TCM service" in our country, and let the Chinese medicine industry go out of the country and into the world to contribute to human health.
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Objective To predict and analyze the structure and function of Stenotrophomonas maltophilia OmpA. Methods Bioinformatics software and biological databases were used to analyze the physicochemical properties, signal peptides,and transmembrane structures of the OmpA protein and to predict the subcellular localization, secondary and tertiary structures,sequence homology and conserved domains,and epitopes of OmpA.Results OmpA protein had strong hydrophilicity but without transmembrane helices in mature protein,and positions 1-22 of the sequence were predicted as signal peptide.In the second structure random coil helix,αlpha-helix,beta-turn and extended strand made up 50.27%, 24.59%,9.29%and 15.85%,respectively.The three-dimensional structure was β-barrel.OmpA was highly conserved among S.maltophilia strains but shared minimal homology with human and mouse proteins.The N-terminal domain of OmpA was OM-channels superfamily and the C-terminal domain of OmpA was OmpA_C-like superfamily.OmpA protein contained 11 dominant antigen epitopes.Conclusion The characteristics of OmpA identified by bioinformatics analysis can not only provide reference for the study of the structure and novel function of OmpA,but also theoretically contribute to the research on related new subunit vaccines.
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Objective:To explore the effect of sulforaphane (SFN) preconditioning on the cold myocardial ischemia-reperfusion injury (IRI) in the rats through PI3K/Akt signaling pathway.Methods:Sixty-four health male Sprague-Dawley (SD) rats were randomly divided into cold IRI group,SFN group,LY (LY294002) + cold IRI group,and LY+SFN group (n=16).The allogeneic heterotopic heart transplantation model was established by donor heart into recipient abdomen.The myocardium tissue was taken 24 h after reperfusion for the detection of histological changes using HE staining.The expression levels of Akt,p-Akt,Bax and Bcl-2 proteins were detected by immunohistochemistry and Western boltting methods.Results:The morphological results showed that the myocardium tissue damage was serious in cold IRI group and LY+cold IRI group,it was light in SFN group;the myocardium tissue damage of the rats in SFN+ LY group was ranged between cold IRI group and SFN group.Compared with IRI group,the expression levels of p-Akt protein and Bcl-2 protein in SFN group were increased (P＜0.05),and the expression level of Bax protein was decreased (P＜0.05).After treatment of blockage LY294002,compared with LY-+-cold IRI group,the expression level of p-Akt protein in LY-+-SFN group was not statistically significant (P＞0.05),the expression level of Bcl2 protein was increased (P＜0.05),the expression levels of Bax protein was decreased),and the ratio of Bcl-2/Bax was also increased (P＜0.05).Conclusion:SFN may attenuate cold IRI of heart transplantation through PI3K/Akt signaling pathway in the rats.