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Objective To establish a genetic monitoring method for laboratory quails.Methods Quail microsatellite loci were searched in the literature,and microsatellite DNA loci suitable for quails were screened by an interspecific transfer method in closely related species,namely chickens and ducks.Quail liver DNA was extracted as a template,and the corresponding loci were screened by PCR amplification and agarose gel electrophoresis.On the basis of amplification of the selected microsatellite loci,the number of alleles,polymorphisms,and microsatellite loci combinations for quail genetic quality detection were selected and detection method were developed.Results We preliminary determined 23 microsatellite loci for genetic monitoring of closed-colony laboratory quails.Conclusions A genetic monitoring method for laboratory quails was preliminary developed.
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Molecular knowledge of human gastric corpus epithelium remains incomplete. Here, by integrated analyses using single-cell RNA sequencing (scRNA-seq), spatial transcriptomics, and single-cell assay for transposase accessible chromatin sequencing (scATAC-seq) techniques, we uncovered the spatially resolved expression landscape and gene-regulatory network of human gastric corpus epithelium. Specifically, we identified a stem/progenitor cell population in the isthmus of human gastric corpus, where EGF and WNT signaling pathways were activated. Meanwhile, LGR4, but not LGR5, was responsible for the activation of WNT signaling pathway. Importantly, FABP5 and NME1 were identified and validated as crucial for both normal gastric stem/progenitor cells and gastric cancer cells. Finally, we explored the epigenetic regulation of critical genes for gastric corpus epithelium at chromatin state level, and identified several important cell-type-specific transcription factors. In summary, our work provides novel insights to systematically understand the cellular diversity and homeostasis of human gastric corpus epithelium in vivo.
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Humans , Epigenesis, Genetic , Gastric Mucosa/metabolism , Chromatin/metabolism , Stem Cells , Epithelium/metabolism , Fatty Acid-Binding Proteins/metabolismالملخص
Objective To compare the preparation efficiency of mouse pox and mouse hepatitis antibodies between two substrains of BALB/c and Big-BALB/c (B-BALB/c) mice, and to provide a theoretical basis and reference for the selection of laboratory animals in the preparation of monoclonal antibodies inducedin vivo through hybridoma.Methods Individuals weighing more than 5% of the weight of normal animals at 4 weeks of age (the criterion for late selection is more than 10%) were selected from a population of conventionally bred BALB/c mice and bred individually, and a subline of B-BALB/c mice was prepared after 10 generations of selection. A total of 40 BALB/c mice and 40 B-BALB/c mice aged 10 to 11 weeks, half male and half female, were selected and inoculated with the mousepox monoclonal antibody hybridoma cell line G23 or the murine hepatitis monoclonal antibody hybridoma cell line Y15 pre-treated with liquid paraffin, respectively. Mice ascites containing monoclonal antibodies were obtained by in vivo induction. The antibody titer was tested by indirect ELISA. The mice were grouped based on the sub-strains, gender and inoculation type of hybridoma to analyze the ascites production, antibody titer and antibody production, and to evaluate the antibody preparation efficiency of the two BALB/c mouse sub-strains.ResultsAfter 10 generations of breeding, the body weight of 10-week-old male and female B-BALB/c mice increased by 22.3% and 12.8%, respectively, compared with BALB/c mice of the same age. Compared with BALB/c mice, B-BALB/c mice had better tolerance and adaptation to secondary ascites collection. Compared with BALB/c mice, the ascites production and antibody titer during the preparation of antibodies in B-BALB/c mice were significantly increased, especially in the hybridoma cell line G23 vaccination group (both P＜0.000 1) . After inoculation with the hybridoma cell lines G23 or Y15, the average antibody production of B-BALB/c mice (14.99×104 U and 33.22×104 U) was higher than that of BALB/c mice (5.33×104 U and 19.31×104 U) (both P＜0.01). After inoculation with hybridoma cell line G23, the average antibody production per unit body weight of B-BALB/c mice (0.55×104 U/g) was higher than that of BALB/c mice (0.23×104 U/g) (P＜0.000 1). And the antibody production per unit body weight of female B-BALB/c or BALB/c mice was higher than that of male B-BALB/c or BALB/c mice (bothP＜0.01).Conclusion B-BALB/c mice can be used as an alternative to BALB/c mice in the in vivo induction of monoclonal antibody preparation, which can achieve the purpose of reducing the number of experimental animals used, lowering the labor cost, and improving the efficiency of antibody preparation.
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Objective To investigate the prognosis relevant factors of laryngeal squamous cell carcinoma (LSCC).Methods From January 2013 to December 2013,the clinical data of 71 patients with LSCC who were initially treated in Zhejiang Cancer Hospital were retrospectively analyzed.Results Univariate analysis showed that there were statistically significant differences in survival rate between the group of supraglottic type and glottic type (60.0% vs.86.3% ,χ2 =6.284,P<0.05),the group of N0 and N+(41.7% vs.86.4% ,χ2 =16.803,P<0.01), the group of early and late stage(93.6% vs.50.0% ,χ2 =19.854,P<0.01).There were no statistically significant differences in survival rate between the group of age ≤50,>50-60,>60-70 and >70(88.9% vs.88.2% vs. 79.3% vs.62.5% ,χ2 =3.909,P>0.05),the group of T1+T2 and T3+T4(83.6% vs.62.5% ,χ2 =3.623,P>0.05),the group of high,medium,low differentiated and unsigned(75.0% vs.69.7% vs.83.3% vs.91.7% ,χ2 =3.780,P>0.05),the group of surgery,radiotherapy and surgery+radiotherapy (74.3% vs.90.9% vs.71.4% , χ2 =2.437,P>0.05).Multivariate analysis showed that age( P =0.003),treatment( P =0.048) had significant effect on the prognosis of patients,but tumor location(P=0.766),T stage(P=0.677),N stage(P=0.482),clinical stage(P=0.825),the degree of pathological differentiation(P=0.206) had no significant effect on the prognosis of patients.Conclusion More aggressive treatment should be supplied for patients with N+,advanced clinical stage and age whom the prognosis are usually poor. In addition, the proportion of tracheal tube extraction should be appreciated.
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Objective To investigate the predictive factors for central lymph node metastasis (CLNM) in papillary thyroid microcarcinoma(PTMC).Methods A retrospective study including 2511 cases of PTMC admitted from Jan.2013 to Jan.2016 were enrolled in our study.Chi-square test was used in univariate analysis.Logistic regression analysis was applied for multivariate analysis.The relation between age,sex,tumor size,multifocality,thyroidal extension,nodular goiter and central lymph node metastasis is analyzed.Results Univariate analysis showed that age,sex,tumor size,multifocality,thyroidal extension and nodular goiter had statistical significance.Age less than 45(P＜0.001,x2=17.442);Male gender(P＜0.001,x2=17.029);Tumor size less than 5 mm (P＜0.001,x2=70.164.);Extrathyroid extension factor (P＜0.001,x2=63.197);Nodular hyperplasia factor (P=0.017,x2=5.611).Multivariate analysis showed there was a significant relationship between multifocality and the central lymph node positivity.The odds ratio (OR) was 1.587 in patients with tumor foci ≥2(P＜0.001).While OR increased sharply near to 3 in patients withtumor foci ≥3(OR=2.730).Tumor size(OR=1.926);Extrathyroid extension(OR=1.606).Conclusions Multifocalty,tumor size and thyroidal extension are the main predicative factors for central lymph node metastasis in PTMC.Among them,tumor foci ≥ 3 is an important predictor.Besides the conventional factors such as tumor size,thyroidal extension etc,multifocalty should also be taken into consideration.
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Objective To investigate the clinical features of papillary thyroid microcarcinoma(PTMC) and the risk factors of central lymph node metastasis (CLNM) in PTMC patients.Methods The clinical data of 855 patients with PTMC who underwent prophylactic central lymph node dissection were retrospectively reviewed.Results In the present study,the rate of CLNM was 21.9％ (187/855) in PTMC patients.In the univariate analysis,the incidence of CLNM was significantly higher in male patients,age ＜ 45 years,tumor size ＞ 5 mm,capsular invasion,multiple tumor and intracapsular spread (31.4 ％ vs.19.5 ％,x2 =11.429,P =0.001;29.3 ％ vs.15.9 ％,x2 =22.416,P =0.000;30.0％ vs.14.3％,x2 =30.669,P=0.000;29.4％ vs.16.8％,x2 =19.233,P =0.000;30.0％ vs.19.9％,x2 =8.205,P =0.004;42.9％ vs.21.3 ％,x2 =5.549,P =0.018,respectively).Multivariate analysis showed that male gender,age ＜ 45 years,tumor size ＞ 5mm,multiplicity and capsular invasion were independent risk factors for CLNM in PTMC patients (95％ CI:0.380-0.834,0.349-0.693,1.450-3.060,1.078-2.229,1.024-2.373,respectively).Conclusion A routine prophylactic central lymph node dissection should be considered particularly in male PTMC patients with age ＜45 years,tumor size ＞5 mm,capsular invasion and tumor multiplicity.
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AIM:To study the microRNA profiling in the serum of insulin-resistant mice and the mechanism of insulin resistance induced by related microRNAs.METHODS:A high-fat diet was used to induce insulin resistance model in KM mice.The microRNA profiling in serum of insulin-resistant and normal mice was analyzed by microarray chip and were validated by real-time PCR.miRanda data base was used to forecast target genes.miRBase was used to obtain the se-quences of related microRNAs, based on which protein interactions were predicted using the online analytical tool STRING. RESULTS:In serum of insulin-resistant mice, the expression of miR-125, miR-126, miR-143, miR-30a, miR-199a, miR-127, miR-184, miR-30e, miR-134, miR-195, miR-206, miR-429, miR-212, miR-362, miR-382, miR-154 and miR-466h was significantly up-regulated.miR-211, miR-504, miR-877 and miR-1930 were significantly down-regulated. miR-143 associated with insulin resistance was able to bind to 3'-UTR of fat mass and obesity-associated protein (FTO), and FTO was found to interact with Rpgrip1l, Tmem18, Mc4r, Npy, Hhex, Tcf712, Cdkal1, Slc30a8, Igf2bp2 and Tha-da.CONCLUSION:Twenty-one microRNAs in the serum of insulin-resistant mice induced by a high-fat diet are signifi-cantly different from those of normal mice, in which 17 kinds were significantly up-regulated.miR-143 closely related to in-sulin resistance is able to regulate FTO protein expression, which interacts with other 10 proteins associated with the occur-rence and development of diabetes.The results are also useful for further study of the molecular mechanisms in insulin re-sistance.
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The aim of this investigation was to assess the in-vitro interaction of two antifungal agents, econazole-nitrate and chelerythrine, against ten fluconazole-resistant clinical isolates and one ATCC type strain 10231 of Candida albicans. The checkerboard microdilution method was performed according to the recommendations of the National Committee for Clinical Laboratory Standards, and the results were determined by visual examination. The interaction intensity was tested in all isolates using the fractional inhibitory concentration index [FICI]. These experiments showed synergism between econazole-nitrate and chelerythrine in antifungal activity against C. albicans, and no antagonistic activity was observed in any of the strains tested. Moreover, time-kill curves were performed with selected strains to confirm the positive interactions. The similarity between the results of the FICI values and the time-kill curves revealed that chelerythrine greatly enhances the antifungal effects of econazole-nitrate against isolates of C. albicans. This synergistic effect may markedly reduce the dose of econazole-nitrate required to treat candidiasis, thereby decreasing the econazole-nitrate toxic side effects. This novel synergism might provide a potential combination treatment against fungal infections
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Objective To investigate the molecular mechanisms of protective effects of thioredoxin (Trx) on human vascular endothelial cells in atherosclerosis.Methods The cell models of Trx-overexpressing cells (Ad Trx) and the control cells (Ad-con) were established by adenovirus vector gene transfer technology in human umbilical vein endothelial cells (HUVECs).The oxidized low density lipoprotein,a risk factor of atherosclerosis,was used as a stimulator.Western blot and indirect immunofluorescence were used to detect the protein expression levels and the cellular localization of Trx,adhesion molecules (ICAM-1,VCAM-1) and the upstream signal pathways.Trx activity was detected by insulin disulfide reduction assay,and cellular reactive oxygen species (ROS)production was detected by fluorescent probe DCFH-DA.Results As compared with control group,Trx protein expression level was enhanced in Ad-trx group and the Trx activity in Ad-Trx group was upregulated by (26.2 ±3.3)％.The result of ROS detection showed that overexpression of Trx significantly inhibited the cellular ROS generation.As compared with control group,overexpression of Trx obviously inhibited the adhesion molecules expression but markedly promoted the phosphorylation of Smad3 in endothelial cells with or without oxLDL stimulation (P＜0.05).Pretreatment of cells with SIS3,a specific inhibitor of Smad3 phosphorylation,reversed Trx-induced inhibition of adhesion molecules expression.Further studies showed that pretreatment of cells with SIS3 enhanced oxLDL-induced AP-1 subunit c-fos nuclear expression.Conclusions The enhancement of Smad3 phosphorylation and c-Fos nuclear expression are mainly responsible for the Trx-induced downregulation of adhesion molecules.
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ObjectiveTo evaluate central compartment lymph node metastasis in cN0 papillary thyroid carcinoma(PTC) patients. MethodsThe clinical data of 166 cN0 PTC patients was retrospectively analyzed. ResultsThe central compartment lymph node metastasis rate of age ＜ 45 years group was higher than the group of age≥45 years(39％ vs.22％,P ＜0.05).Tumor size ＞0.5 cm tends to metastasize more often than tumor size ≤0.5 cm(P ＜0.05) ; there was significantly difference in central compartment lymph node metastasis rate between upper pole tumor ( 25％,12/48 ) and middle ( 45％,13/29 ) or lower pole (43％,31/72),P ＜ 0.01. ConclusionscN0 PTC patients with tumor located in middle pole or lower pole,tumor size ＞ 0.5 cm or age ＜ 45 years are at higher risks for central compartment lymph node metastasis.
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OBJECTIVE@#To discuss the effectiveness and advantage of application of pectoralis major flap in the reconstruction of massive defects after head and neck tumor excision.@*METHOD@#Thirteen patients with head and neck carcinoma underwent tumor excision, and the massive defects were fixed by pectoralis major myocutaneous flap. Ten patients with recurrent laryngeal carcinoma and 2 with hypopharyngeal carcinoma and 1 with thyroid carcinoma were included.@*RESULT@#Eleven cases healed well, 1 case appearance fistula of parotid gland, 1 case was lost to follow-up.@*CONCLUSION@#Pectoralis major myocutaneous flap with adequate vascularization, enough tissue and ideal healing ability cab be used to reconstruct massive defects. And so it can play an important role in providing a better life expectancy and can radically cure or relieve the complaint.
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Adult , Aged , Female , Humans , Male , Middle Aged , Head and Neck Neoplasms , General Surgery , Pectoralis Muscles , Transplantation , Postoperative Period , Plastic Surgery Procedures , Methods , Surgical Flapsالملخص
0.05). CONCLUSION The secar of oropharynx can be regarded as a predictive index for OSAHS severity and guideline for operation, but the change of secar after operation can not be used as a predicitive index for therapeutic effectiveness.
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OBJECTIVE@#To study the pathologic diagnosis and the injury time estimation in light closed encephalon injury.@*METHODS@#Mice were hurt by fluid percussion, and were killed at 15, 30 min, 1, 3 , 6, 12 h, 1, 4, 7, 14 d respectively after injury. The expression of Fas-L in the cerebral cortex, thalamus, and hippocampi was detected by immunohistochemistry and the results were assessed by image analysis system.@*RESULTS@#It is showed that the expression of Fas-L could be detected in 1 h after injury, and increased significantly in three hours, and it reached apex 12 h after injury, and decreased gradually four days after injury, and returned normal 14 days after injury.@*CONCLUSION@#This research demonstrated that Fas-L mediated apoptosis appeared not only around brain trauma but also in the brain tissue far away from the traumatic area. It indicted that the expression of Fas-L is a useful target for diagnosis of early brain injury; the regularity of Fas-L expression could be used as one of indication to date the time of brain injury.
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Animals , Male , Rats , Apoptosis , Brain/metabolism , Brain Injuries/pathology , Fas Ligand Protein , Image Processing, Computer-Assisted , Immunohistochemistry , Membrane Glycoproteins/biosynthesis , Rats, Wistar , Time Factors , Tumor Necrosis Factors/biosynthesisالملخص
Whether the ATP-sensitive potassium channel opener pinacidil can provide myocardial protective effects in prolonged isolated global ischemic rat heart was investigated. On modified isolated rat working heart model, 40 hearts were divided into four groups randomly: Hyperpolarized arrest H-K solution containing pinacidil (50 mumol/L) (P1 and P2) and depolarized arrest St. Thomas' solution (S1 and S2) subjected to 15 degrees C hypothermia, 60 min (P1 and S1) or 120 min (P1 and S2) of ischemia and 30 min reperfusion. The experimental indices included cardioplegic efficiency, cardiac function, coronary blood flow, myocardial enzyme release, myocardial water and ATP content. Hyperpolarized arrest provided significantly better recovery of cardiac function than depolarized arrest. Postischemic coronary flow and myocardial ATP content were higher. The arrest time of electro-mechanical activities were longer than depolarized arrest. There were no differences among the groups in myocardial water contents. The hyperpolarized arrest solution containing pinacidil can provide a marked myocardial protective effect during prolonged hypothermic myocardial ischemia.
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Animals , Male , Rats , Cardioplegic Solutions , Pharmacology , Cyclic Nucleotide-Gated Cation Channels , Heart Arrest, Induced , Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels , In Vitro Techniques , Ion Channels , Metabolism , Myocardial Reperfusion Injury , Myocardium , Metabolism , Pinacidil , Pharmacology , Potassium Channels , Random Allocation , Rats, Wistarالملخص
AIM: To explore whether AT2 receptor is expressed in skeletal muscular vasculature, and the expression of angiotensin Ⅱ receptors in pulmonary circulation of children with left-to-right shunt but without obstructive pulmonary hypertension. METHODS: Lung and skeletal muscular tissue were obtained from 20 children with left-to-right shunt by biopsy during operation. These skeletal muscular tissues were detected by reverse transcriptase/polymerase chain reaction (RT-PCR ) and immunohistochemistry techniques for AT2 receptors. mRNA of AT1 and AT2 receptors in lung tissues were detected by RT-PCR and analyzed semi-quantitatively. RESULTS: In all of the skeletal muscular tissues, mRNA of AT2 receptor was found by RT-PCR, and the results of immunohistochemistry staining for AT2 receptor of vessels were positive. In the lung tissues, the level of mRNA of AT2 receptor was different to AT1 receptor, and the former was higher than the latter (P
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Objective To explore the effect of estradiol benzoate on the expressions of heat shock protein (HSP)70, 90 in endometrial glandular epithelial cell. Methods Normal endometrial glandular epithelial cells were isolated, and cultured by enzymolysis method and identified by electron microscopy and immunohistochemical analysis. The normal endometrial glandular epithelial cells were treated with culture medium only, estradiol benzoate (10 -9 , 10 -8 , 10 -7 or 10 -6 mol/L), estradiol benzoate(10 -9 , 10 -8 , 10 -7 or 10 -6 mol/L) and antiestrogen ICI 182780(fulvestrant,faslodex, 1?mol/L)and ICI 182780 only for 6, 12, 18, 24 hours respectively. The dose-and time-related effect of estradiol benzoate and ICI 182780 on the cell growth was measured by mononuclear cell direct cytotoxicity assay (methyl thiazolyl tetrazolium assay), and that on the expression of HSP70 and HSP90 in normal endometrial glandular epithelial cell in vitro was measured by immunohistochemical analysis and computerized image analysis system. Results Estradiol benzoate stimulated cell growth in a time-and dose-dependent manner and the effect was attenuated by the antiestrogen ICI 182780. The average cell growth rates of 10 -9 , 10 -8 , 10 -7 , 10 -6 mol/L estradiol benzoate for 24 hours were(170?9)%,(207?11)%,(231?12)%,(257?10)%, which were significantly higher than those of 6 hours (117?13)%, (129?10)%, (146?10)%, (176?6)%, P