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ObjectiveTo screen out the transcriptomes related to the intervention of Wuzi Yanzongwan on the spermatogenic function of semi-castrated male mice, and to explore its potential mechanism in the intervention of the progress of low spermatogenic function. MethodBalb/c mice were randomly divided into sham-operated group, model group, testosterone propionate group(0.2 mg·kg-1·d-1, intramuscular injection) and Wuzi Yanzongwan group(1.56 g·kg-1·d-1, intragastric administration) according to body weight, with 12 mice in each group. The right testicle and epididymis were extracted from the model group and the drug administration group to construct the semi-castrated model of low spermatogenic function, while the fur and the right scrotum of the sham-operated group were only cut and immediately sterilized and sutured. At the end of the intervention, hematoxylin-eosin(HE) staining was used to observe the histopathology of testis, enzyme-linked immunosorbent assay(ELISA) was used to detect the levels of serum testosterone(T), luteinizing hormone(LH) and follicle stimulating hormone(FSH). The sperm count and motility of epididymis were measured by automatic sperm detector of small animal. Transcriptomic microarray technology was used to detect the mRNA expression level of testicular tissue in each group, the transcriptome of genes related to the regulation of Wuzi Yanzongwan was screened, and three mRNAs were selected for Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) to verify the transcriptome data. Through the annotation analysis of Gene Ontology(GO) and the signaling pathway analysis of Kyoto Encyclopedia of Genes and Genomes(KEGG), the related functions of drugs regulating transcriptome were analyzed. ResultCompared with the sham-operated group, the testicular tissue of mice in the model group showed spermatogenic injury, contraction and vacuolization of the seminiferous tubules, reduction of spermatogenic cells at all levels, widening of the interstitial space, obstruction of spermatogonial cell development and other morphological abnormalities, and serum T significantly decreased, LH significantly increased(P<0.01), and FSH elevated but no statistically significant difference, the count and vitality of epididymal sperm significantly decreased(P<0.01). There were 882 differentially expressed mRNAs in the testicular tissues, of which 565 were up-regulated and 317 were down-regulated. Cluster analysis showed that these differentially expressed mRNA could effectively distinguish between the sham-operated group and the model group. Compared with the model group, the damage to testicular tissue in the Wuzi Yanzongwan group was reduced, the structure of the seminiferous tubules was intact, vacuolization was reduced, and the number of spermatogenic cells at all levels was significantly increased and arranged tightly. The serum T significantly increased, LH significantly decreased(P<0.01), and FSH decreased but the difference was not statistically significant. The count and vitality of sperm in the epididymis were significantly increased(P<0.01). Moreover, Wuzi Yanzongwan could regulate 159 mRNA levels in the testes of semi-castrated mice, of which 32 were up-regulated and 127 were down-regulated, and the data of the transcriptome assay was verified to be reliable by Real-time PCR. GO and KEGG analysis showed that the transcriptome functions regulated by Wuzi Yanzongwan were involved in the whole cell cycle process of sperm development such as sex hormone production of interstitial cells in testis, renewal, differentiation, metabolism, apoptosis and signal transduction of spermatogenic cells, and were closely related to the biological behaviors of signaling pathways such as spermatogenic stem cell function, endoplasmic reticulum protein processing and metabolic program. ConclusionWuzi Yanzongwan can effectively improve the low spermatogenic function of semi-castrated male mice, and its mechanism may be related to the regulation of testicular transcriptional regulatory network, the synthesis of sex hormones in testicular interstitial cells, the function of spermatogenic stem cells, the whole cell cycle process of spermatogenesis, as well as the expression of endoplasmic reticulum protein processing and metabolic program related genes transcription.
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Objective:To evaluate the accuracy of ultrasonography in determining the laryngeal mask airway position using three-dimensional CT reconstruction (3D-CTR) images as the standard.Methods:One hundred and thirty-four patients, aged 18-64 yr, weighing 30-100 kg, of American Society of Anesthesiologists physical status Ⅰ or Ⅱ, scheduled for elective cerebral vascular interventional stent surgery in our hospital, were selected.Cervical ultrasonography was performed before and after laryngeal mask airway placement, and digital subtraction angiography was performed during operation to obtain 3D-CTR images.Neck ultrasound and 3D-CTR images were analyzed and graded.Results:A total of 125 patients completed the study.At the vertical position of the laryngeal mask airway, ultrasonic grading was positively correlated to 3D-CTR grading ( r=0.742, P<0.05). The sensitivity, specificity, positive predictive value, negative predictive value and accuracy of ultrasound in determining the rotation of the laryngeal mask airway was 73.0% (95% CI: 62.0%-84.4%), 92.8% (95% CI: 86.9%-98.7%), 89.1% (95% CI: 80.5%-97.7%), 81.0% (95% CI: 72.6%-89.4%), and 74.2% (95% CI 65.2%-81.1%), respectively.In terms of laryngeal mask airway depth grading, the correlation between ultrasonic grading was positively correlated to 3D-CT grading ( r=0.332, P<0.05). The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of ultrasound in determining the depth of the laryngeal mask airway was 58.5% (95% CI: 46.9%-70.1%), 93.3% (95% CI: 87.2%-99.4%), and 90.5% (95% CI: 82.0%-99.0%), 67.5% (95% CI: 57.7%-77.3%), and 33.2% (95% CI: 16.8%-47.8%), respectively.The positive predictive value of ultrasound in determining the optimal placement of the laryngeal mask airway was 61.1% (95% CI: 48.6%-73.6%). Conclusion:Although ultrasound cannot measure the depth of the tip of the laryngeal mask airway into the esophagus, it has higher accuracy in determining the rotation and can be used as a reliable tool for clinical detection of the position of the laryngeal mask airway, especially the rotation.
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Objective:To analyze the long-term efficacy and safety of re-irradiation for recurrent glioma.Methods:The data of 52 patients with recurrent gliomas were collected from 2009 to 2019. The median planned targetvolume (PTV) was 73.5 cm 3(49.9-102.7 cm 3) and the median dose was 45.0 Gy (43.0-48.8 Gy). Kaplan-Meier method was used for survival assessment, log-rank test for difference assessment, and Cox’s regression model for multivariate prognostic analysis. Results:The median follow-up time was 32.6 months. The median overall survival (OS) and progression-free survival (PFS) time were 16.1 months (95% CI, 4.1-28.1) and 8.0 months (95% CI, 4.0-12.0). The 1-, 2-and 3-year survival rates were 67%, 43% and 29%, respectively. The 6-month, 1-year and 2-year PFS rates were 67%, 40%, 26%, respectively. Multivariate analysis showed that KPS score and recurrence time significantly affected the OS ( P=0.012, P=0.001). KPS score and time interval between two radiotherapies significantly impacted the PFS ( P=0.003, P=0.018). Stratified analysis showed that KPS score was the independent prognostic factor of OS and PFS in patients with WHO grade Ⅱ initial pathology and reoperation after recurrence ( P<0.001, P=0.012); clinical manifestation was the independent prognostic factor of OS and PFS in patients with WHO grade Ⅲ and Ⅳ initial pathology ( P=0.006, P=0.044). The overall incidence of adverse reactions was 30.8%. Grade 1 adverse reactions accounted for 25.0%, and 5.8% for grade 2. Conclusions:Re-irradiation for recurrent glioma yields good long-term clinical efficacy and tolerable adverse reactions.
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Objective To analyze the association between polycyclic aromatic hydrocarbons(PAHs)exposure and rheumatoid arthritis (RA)based on large sample data.Methods The RA patients(RA group)and non-RA patients(non-RA group)with complete data were selected from the National Health and Nutrition Survey Database in the United States(NHANES)(2005-2014).The logistic regression model was used to analyze the association between 8 monohydroxylated(OH-)PAH metabolites in the urine and RA.Results A total of 357 RA patients and 5,256 non-RA participants were included.After adjusting the confounding factors by logistic analysis,the level of OH-PAHs mixture at the highest quartile(Q4)was associated with increased risk of RA compared with that at the lowest quartile(Q1) (OR =1.60,95 % CI:1.16 2.23).For a single kind of OH-PAHs,the Q4 levels of 1-hydroxynaphthalene (OR =1.59,95 % CI.1.14-2.23),2-hydroxynaphthalene (OR =1.66,95 % CI:1.19-2.32),2-hydroxyfluorene(OR =1.61,95 % CI:1.17-2.22),3-hydroxyfluorene(OR =1.64,95% CI:1.18-2.27) and 1-hydroxyphenanthrene (OR =1.38,95 % CI:1.00-1.94) were all associated with significantly increased risk of RA compared with the Q1 level(all P<0.05).However,the Q2 level of 1-hydroxypyrene(OR =0.60,95% CI:0.43-0.83) was related to a decreased incidence of RA (P<0.01).Conclusions OH-PAHs mixed exposure is a risk factor for RA.The association between the level of individual OH-PAH and the rate of RA is bidirectional and is depended on the type and concentration of OH-PAHs.
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Objective@#To analyze the association between polycyclic aromatic hydrocarbons(PAHs)exposure and rheumatoid arthritis(RA)based on large sample data.@*Methods@#The RA patients(RA group)and non-RA patients(non-RA group)with complete data were selected from the National Health and Nutrition Survey Database in the United States(NHANES)(2005—2014). The logistic regression model was used to analyze the association between 8 monohydroxylated(OH-)PAH metabolites in the urine and RA.@*Results@#A total of 357 RA patients and 5, 256 non-RA participants were included.After adjusting the confounding factors by logistic analysis, the level of OH-PAHs mixture at the highest quartile(Q4)was associated with increased risk of RA compared with that at the lowest quartile(Q1)(OR=1.60, 95%CI: 1.16-2.23). For a single kind of OH-PAHs, the Q4 levels of 1-hydroxynaphthalene(OR=1.59, 95%CI: 1.14-2.23), 2-hydroxynaphthalene(OR=1.66, 95%CI: 1.19-2.32), 2-hydroxyfluorene(OR=1.61, 95%CI: 1.17-2.22), 3-hydroxyfluorene(OR=1.64, 95%CI: 1.18-2.27)and 1-hydroxyphenanthrene(OR=1.38, 95%CI: 1.00-1.94)were all associated with significantly increased risk of RA compared with the Q1 level(all P<0.05). However, the Q2 level of 1-hydroxypyrene(OR=0.60, 95%CI: 0.43-0.83)was related to a decreased incidence of RA(P<0.01).@*Conclusions@#OH-PAHs mixed exposure is a risk factor for RA.The association between the level of individual OH-PAH and the rate of RA is bidirectional and is depended on the type and concentration of OH-PAHs.
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<b>Objective::To study the chemical constituents of pericarps of <italic>Zanthoxylum bungeanum</italic>. <b>Method::The dried pericarps of <italic>Z</italic>. <italic>bungeanum</italic> were smashed, and then extracted and concentrated in 95%ethanol to obtain the total extract. The total extract was loaded on a silica gel CC, eluted with different polar solvents in sequence, and then concentrated to obtain corresponding parts. The methylene chloride fraction and the <italic>n</italic>-butanol fraction were separated and purified by silica gel column chromatography, Sephadex LH-20 gel column chromatography, ODS column chromatography, semi-preparative HPLC, etc. And their structures were identified based on physicochemical properties and various spectroscopic methods. <b>Result::Fourteen compounds were isolated from the dichloromethane fraction and the n-butanol fraction of the <italic>Z</italic>. <italic>bungeanum</italic> and identified as(1<italic>S</italic>, 3<italic>S</italic>)-1-methyl-1, 2, 3, 4-tetrahydro-<italic>β</italic>-carboline-3-carboxylic acid(<bold>1</bold>), (3<italic>S</italic>)-1, 2, 3, 4-tetrahydro-<italic>β</italic>-carboline-3-carboxylic acid(<bold>2</bold>), apigenin-7, 4′-dimethyl ether(<bold>3</bold>), genkwanin(<bold>4</bold>), lcariside F<sub>2</sub>(<bold>5</bold>), breyniaionoside A(<bold>6</bold>), 3-methoxyphenethyl alcohol-4-<italic>O</italic>-<italic>β</italic>-<italic>D</italic>-glucopynanoside(<bold>7</bold>), 1-<italic>O</italic>-<italic>β</italic>-<italic>D</italic>-glucopyranosyloxy-3-methoxy-5-hydroxybenzene(<bold>8</bold>), orcinol-<italic>O</italic>-<italic>β</italic>-<italic>D</italic>-glucopyranoside(<bold>9</bold>), syringin(<bold>10</bold>), 4-[(3<italic>S</italic>)-3-hydroxybutyl]-2-methoxyphenyl-<italic>β</italic>-<italic>D</italic>-glucopyranoside(<bold>11</bold>), (+ )-lyoniresinol-3a-<italic>O</italic>-<italic>β</italic>-<italic>D</italic>-glucopyranoside(<bold>12</bold>), 2-methylpropanyl-6-<italic>O</italic>-<italic>β</italic>-<italic>D</italic>-apiofuranosyl-<italic>β</italic>-<italic>D</italic>-glucopyranoside(<bold>13</bold>)and(<italic>E</italic>)-6-hydroxy-2, 6-dimethylocta-2, 7-dienoic acid(<bold>14</bold>). <b>Conclusion::All compounds were isolated from <italic>Z</italic>. <italic>bungeanum</italic> for the first time, and compounds <bold>1-4, 12</bold> and <bold>14</bold> were isolated from this genus for the first time.
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Objective To investigate PLCE1 protein expression in esophageal squamous cell carcinoma (ESCC)tissues,and understand the effect of PLCE1 protein expression on the prognosis of patients with ESCC.Methods The PLCE1 protein were detected in 85 patients with surgically resected ESCC paraffin-embedded tissue using immunohistochemical staining,the patients' information were selected from March 1997 to December 2011 in the database of the Henan Key Laboratory for Esophageal Cancer Research,and the survival analysis were operated with Kaplan Meier single factor and Cox multivariate regression.Results In the 85 cases,the lost to follow-up were 5 cases,the follow-up rate was 94.1% (80/85).In the 80 cases with ESCC,the positive expression rate of PLCE1 was 77.5% (62/80).PLCE1 positive expression was not significantly associated with differentiation,infiltration depth,lymph node metastasis and family history with ESCC (P > 0.05).The univariate survival analysis by Kaplan-Meier showed that protein expression of PLCE1 had no significant association with overall survival (P > 0.05).Cox regression survival analysis showed that the lymph node metastasis was associated with the survival time of ESCC patients [the relative risk were 1.763,95% CI(1.008,3.084),P =0.047].Conclusion The expression of PLCE1 protein was involved in mechanism of ESCC,and might not predicted the prognosis of the patients with ESCC.
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Objective:To study the differentiation capacity of the fibroblast-like cells isolated from human skin dermis into mesenchymal stem cells, and to explore the feasibility to use these cells as alternative cell source of autologus bone marrow mesenchymal stem cells (BMSCs ) for regeneration of tissue inj uries and defects. Methods:Full thickness skin samples were obtained from the abdomen of surgical patients, then digested with dispase and collagenase Ⅰ subsequently. Thereafter, the digested cells were collected and cultured, followed by suspension with serum free medium containing N2,B27,basic fibroblast growth factor (bFGF),and epidermal growth factor (EGF).The skin dermis derived spheroids (SDDSs)were collected and monolayer cultured in serum-containing medium.Finally,the cells were characterized by immunofluorescence staining and differentiation assays.Results:The dermis derived cells proliferated and formed SDDSs in the suspension of serum-free medium. After monolayer cultivation in serum-containing medium, the cells from spheroids were successfully expanded to large number. The cells expressed mesenchymal stem cells markers CD90, CD105 and vimentin. Under osteogenic,chondrogenic and adipogenic differentiation conditions,these cells were differentiated into the alizarin red,safranin O, and oil red O staining positive cells, displayed similar differentiation traits with BMSCs. However,safranin O staining was weaker in the dermis derived cells than BMSCs. Conclusion:A kind of fibroblast-like cells exist in human skin dermis, and have osteocytic, chondrogenic and adipogenic differentiation potentials,demonstrating that these cells will be utilized as a novel cell source for repairing the tissue injury and defect in clinic.