1.IN VIVO EFFECT OF 3′,5′-cAMP ON EHRLICH ASCITES TUMOR CELLS AND THE CHANGES OF THE INTRACELLULAR 3′,5′_cAMP,cAMP-PDE ACTIVITY AND THE CHANGES OF MICROVILLI IN THESE TUMOR CELLS
Daishu WANG ; Menglian ZHAO ; Zhongxiang LIN ; Yunyan LIANG ; Kunren WANG ;
Acta Anatomica Sinica 1953;0(01):-
This experiment was carried out in young adult albino mice,which were injectedwith Ehrlich ascites tumor cells intraperitoneally to produce Ehrlich ascites tumor.The animals were divided into 2 groups.The first group received cAMP plusaminophylline for 5 to 13 days after inoculation.The second group received saline ascontrol.We found that in the administration of cAMP together with aminophylline,thegrowth of Ehrlich ascites tumor cells was inhibited to the extent of 53% at the 9thdays after inoculation.Early or later than the 9th day,the inhibitory rates were marklylower.By using cAMP immunocytochemical method,it was found that on the9th day of inoculation,there was an increase of the intensity of intracellualr cAMPspecific fluorescence in tumor cells of the cAMP treated mice in comparison withthose of the control.It also inhibited the 3',5'-cAMP-PDE activity on the 5th to 7thday after inoculation.Under the dark field microscope on the surface of the Ehrlich tumor cells therewere numerous“brush-like”microvilli,but they were not visible on the surfaceafter treated with cAMP together with aminophylline.The agglutination by theCon.A,was decreased markly on the 7th to 9th day after the inoculation.The possible relationship between the level of intracellular cAMP and of the3',5'-cAMP-PDE,and especially the inhibition of the formation of microvilli onthe treated tumor cell surface is discussed in this paper.
2.OBSERVATION OF THE MICROFILAMENTS IN THE CULTURED HUMAN FIBROBLASTS WITH COOMASSIE BLUE R 250
Menglian ZHAO ; Minying NIU ; Baorong FAN ; Jie GAO ; Kunren WANG ;
Acta Anatomica Sinica 1954;0(02):-
The experiments was conducted on the fibroblasts of human prepuce in vitro todemonstrate the distribution of cytoplasmic microfilaments with Coomassie Blue R250.It has been found that the distribution of the cytoplasmic microfilaments canbe disturbed by the specific inhibitor,Cytochalasin B.We also observed the inhi-bition by Cytochalasin B was reversible.The cytoplasmic microfilaments regainedtheir normal distribution after the Cytochalasin B was removed.This result further confirmed that the staining method of Coomassie BrilliantBlue R 250 is reliable to demonstrate microfilaments in cultured fibrablasts.
3.VARIATION IN LOCALIZATION OF CONCANAVALIN A RECEPTORS ON GASTRIC CANCER CELL LINE AND FIBROBLASTS
Jiacun FANG ; Minying NIU ; Yongjin SHI ; Daishu WANG ; Menglian ZHAO ;
Acta Anatomica Sinica 1957;0(04):-
The distribution of receptor sites for concanavalin A (Con A) on the cellsurface of human gastric cancer cell line (MGC 80-3) and fibroblast cell cultures wasstudied with electron and light microscope after application of Con A-peroxidasemethod.The cytochemical reaction of the majority of non-prefixed MGC 80-3 cellsshows a striking tendency to be more uneven distribution than on fibroblasts.Clustering,patching and capping of Con A binding sites on the cancer cells anduniform distribution on the fibroblasts were observed.The effects of incubationtemperature,prefix treatment,different phase of cell cycles and cell growth condi-tions on the distribution of Con A receptor complexes were observed.The possiblereasons for the more irregular distribution of the cytochemical reaction product onthe MGC 80-3 cells than the fibroblasts are discussed.
4.Prevalence of depressive symptoms among grassroots healthcare workers in Zhejiang Province
LUO Kemeng ; LI Sheng ; LÜ ; Menglian ; LIN Junfen
Journal of Preventive Medicine 2023;35(10):881-884
Objective:
To investigate the prevalence and influencing factors of depressive symptoms among grassroots healthcare workers in Zhejiang Province, so as to provide insights into improving their mental health.
Methods:
Grassroots healthcare workers of community health service centers and township health centers were sampled from one county (city, district) in each of 11 cities in Zhejiang Province using a convenience sampling method from December 2022 to January 2023. Participants' gender, age, educational level and average daily sleep duration in the past week were collected through questionnaires, and depression symptoms was investigated according to Self-rating Depression Scale. Factors affecting the depressive symptoms were identified using a multivariable logistic regression model.
Results:
A total of 1 946 questionnaires were distributed, and 1 945 valid questionnaires were recovered, with an effective response rate of 99.95%. There were 444 boys, accounting for 22.83%, and 1 501 girls, accounting for 77.17%. The median age was was 36 (interquartile range, 44) years. There were 786 healthcare workers detected with depressive symptoms, with a prevalence rate of 40.41%, and the prevalence rates of mild, moderate and severe depressive symptoms were 26.94%, 10.49% and 2.98%, respectively. Multivariable logistic regression analysis identified age (50 years and older, OR=0.572, 95%CI: 0.386-0.846), annual income (100 000 to 149 999 Yuan, OR=0.780, 95%CI: 0.635-0.958; 150 000 to 199 999 Yuan, OR=0.463, 95%CI: 0.282-0.760; 200 000 Yuan and above, OR=0.303, 95%CI: 0.098-0.937), vocation (nurse, OR=1.593, 95%CI: 1.252-2.027) and sleep duration (less than 7 hours, OR=2.164, 95%CI: 1.768-2.648) as factors affecting depressive symptoms among grassroots healthcare workers in Zhejiang Province.
Conclusions
The prevalence of depressive symptoms among grassroots healthcare workers in Zhejiang Province is 40.41%. Age, annual income, job and sleep duration may affect the development of depressive symptoms among grassroots healthcare workers.
5.Cross-species Transmission of Avian Leukosis Virus Subgroup J.
Yanwei SHEN ; Menglian HE ; Ji ZHANG ; Manda ZHAO ; Guihua WANG ; Ziqiang CHENG
Chinese Journal of Virology 2016;32(1):46-55
Avian leukosis virus subgroup J (ALV-J) is an avian retrovirus that can induce myelocytomas. A high-frequency mutation in gene envelope endows ALV-J with the potential for cross-species transmission. We wished to ascertain if the ALV-J can spread across species under selection pressure in susceptible and resistant hosts. First, we inoculated (in turn) two susceptible host birds (specific pathogen-free (SPF) chickens and turkeys). Then, we inoculated three resistant hosts (pheasants, quails and ducks) to detect the viral shedding, pathologic changes, and genetic evolution of different isolates. We found that pheasants and quails were infected under the selective pressure that accumulates stepwise in different hosts, and that ducks were not infected. Infection rates for SPF chickens and turkeys were 100% (16/16), whereas those for pheasants and quails were 37.5% (6/16) and 11.1% (3/27). Infected hosts showed immune tolerance, and inflammation and tissue damage could be seen in the liver, spleen, kidneys and cardiovascular system. Non-synonymous mutation and synonymous ratio (NS/S) analyses revealed the NS/S in hypervariable region (hr) 2 of pheasants and quails was 2.5. That finding suggested that mutation of isolates in pheasants and quails was induced by selective pressure from the resistant host, and that the hr2 region is a critical domain in cross-species transmission of ALV-J. Sequencing showed that ALV-J isolates from turkeys, pheasants and quails had moved away from the original virus, and were closer to the ALV-J prototype strain HPRS-103. However, the HPRS-103 strain cannot infect pheasants and quails, so further studies are needed.
Amino Acid Sequence
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Animals
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Avian Leukosis
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transmission
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virology
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Avian Leukosis Virus
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classification
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genetics
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physiology
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Chickens
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Ducks
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virology
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Galliformes
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virology
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Host Specificity
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Molecular Sequence Data
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Poultry Diseases
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transmission
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virology
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Quail
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virology
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Sequence Alignment
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Turkeys
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virology
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Viral Envelope Proteins
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chemistry
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genetics
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metabolism
6.PREPARATION OF MONOCLONAL ANTIBODY AGAINST PROTEIN KINASE A AND ITS IMMUNOCYTOCHEMICAL LOCALIZATION IN NORMAL AND MALIGNANT CELLS
Jiachun FANG ; Yongjin SHI ; Minying NIU ; Yunyan LIANG ; Menglian ZHAO ; Daishu WANG
Acta Anatomica Sinica 1957;0(04):-
A rat monoclonal antibody of the IgG1 class, McAb B5A8, specific for the cAMP-dependent protein kinase(PKA) was produced. Western blot analysis revealed specific binding of the antibody to protein of 52-56 kd.The affinity-purified McAb BSA8 was labeled with FITC. Immunofluorescent localization of PKA was examined in human fibroblasts, gastric cancer cell line (MGc 80-3)and EAC cells. The distribution of PKA on the cytoplasmic microtubule network, Golgi region and nucleoli were observed. PKA was localized in the nuclear region in G2 phase of synchronized MGc 80-3 cells, and it was only found around the nuclei of MGc 80-3 cells which were incubated with DBcAMP. The changes in the distribution of PKA in cells are discussed.
7.Establishment and evaluation of a predicative model in diagnosing unilateral primary aldosteronism
Yangjie ZENG ; Fan YANG ; Yijie WANG ; Zhaoyang TIAN ; Menglian LI ; Ping LI ; Dalong ZHU
Chinese Journal of Endocrinology and Metabolism 2023;39(10):870-875
Primary aldosteronism(PA) is a common cause of secondary hypertension, with adrenal vein sampling(AVS)being the gold standard for etiological typing diagnosis. This retrospective study analyzed data from patients diagnosed with PA who underwent AVS or unilateral adrenalectomy at Nanjing Drum Tower Hospital′s Endocrinology Department from July 2018 to August 2021. Using multivariate logistic regression analysis, factors associated with unilateral aldosterone hypersecretion were identified. Based on these factors, a predictive model for diagnosing unilateral primary aldosteronism(UPA)was established: age <40 years, plasma aldosterone concentration(PAC)>15 ng/dL, adrenal CT indicating an unilateral typical adenoma, and spontaneous hypokalemia. This predictive model could help 14% of PA patients avoid unnecessary AVS procedures.
8.The diagnostic significance of tumor diameter and unenhanced CT attenuation value in distinguishing benign from malignant adrenal masses
Menglian LI ; Ziwei ZHANG ; Mengsi LIU ; Yangjie ZENG ; Zhaoyang TIAN ; Fan YANG ; Ping LI
Chinese Journal of Endocrinology and Metabolism 2023;39(12):1016-1022
Objective:To explore the pathological characteristics of adrenal masses based on various tumor diameter and unenhanced computed tomography(CT) attenuation value, and evaluate the value of the two parameters in the assessment of the benign and malignant nature of adrenal masses.Methods:The data of 1 367 patients who underwent adrenalectomy in Nanjing Drum Tower Hospital from January 2017 to October 2022 were retrospectively collected. The adrenal masses were divided into four groups according to tumor diameter and unenhanced CT attenuation value, and the clinical and histopathological characteristics of the four groups were compared respectively. Logistic regression was used to analyze the correlation between tumor diameter, non-contrast CT attenuation value and malignant adrenal masses, and receiver operating characteristic(ROC) curve was used to assess the diagnostic value of both in benign and malignant adrenal masses.Results:The proportion of adrenocortical carcinoma and other malignant tumors increased with the rise of tumor diameter or unenhanced CT attenuation value. After adjusting for age and gender, tumor diameter( OR=1.624, 95% CI 1.464-1.803, P<0.001) and unenhanced CT attenuation value( OR=1.108, 95% CI 1.079-1.138, P<0.001) were predictors of malignant adrenal masses. The tumor diameter and unenhanced CT attenuation value in diagnosing malignant adrenal masses had area under the ROC curve(AUC) of 0.838 and 0.892, respectively. With the optimal cut-off values of >3.4 cm and >30 HU, the sensitivity was 75.5% and 83.7%, and the specificity was 80.5% and 84.4%, respectively. The combination of tumor diameter >3.4 cm and unenhanced CT attenuation value >20 HU had an AUC of 0.927, with a sensitivity of 71.4% and a specificity of 90.1% in diagnosing malignant adrenal masses. Conclusions:Tumor diameter and unenhanced CT attenuation value has important significance in the differential diagnosis of benign and malignant adrenal masses. A combination of tumor diameter (>3.4 cm) and unenhanced CT attenuation value (>20 HU) demonstrates best diagnostic efficiency. Clinical application of this combined index can effectively diagnose malignant adrenal masses while avoiding unnecessary investigations or surgery.
9.Clinical features and prognostic factors of adrenocortical carcinoma with hypercortisolism
Ziwei ZHANG ; Menglian LI ; Chaoyang TIAN ; Yangjie ZENG ; Yijie WANG ; Mengsi LIU ; Ping LI
Chinese Journal of Endocrinology and Metabolism 2023;39(12):1010-1015
Objective:To identify the clinical and pathological characteristics of adrenocortical carcinoma(ACC) with hypercortisolism and analyze the prognostic factors.Methods:Clinical data of ACC patients between January 2003 and December 2022 from Nanjing Drum Tower Hospital were collected retrospectively. Clinical and pathological characteristics were compared between ACC patients with hypercortisolism and nonfunctional ACCs. Kaplan-Meier method was used for survival analyses and Cox regression models were performed to analyze prognostic factors for ACC patients.Results:In 61 cases of ACC patients, the average age was (49.33±16.32) years. After a median follow-up of 77 months(95% CI 47.49-106.51 months), median overall survival and progression-free survival were 50 months(95% CI 20.44-79.56 months) and 29 months(95% CI 22.87-35.13 months), respectively. Among 36 patients with complete endocrinologic evaluations, 19(52.77%) were diagnosed with hypercortisolism. Compared with nonfunctional ACC, patients with hypercortisolism had more hypokalemia(42.11% vs 6.25%, P=0.022), capsular invasion(68.42% vs 25.00%, P=0.018), and distant metastases(73.68% vs 25.00%, P=0.007). Median overall survival and progression-free survival were significantly shorter than nonfunctional ACC(overall survival: 39 months vs 67 months, P=0.009; progression-free survival: 30 months vs 51 months, P=0.040) as well. Multivariate Cox regression analyses indicated that Ki67 index( HR=1.078, P=0.024) was an independent risk factor for overall survival. Hypercortisolism( HR=71.112, P=0.006), Ki67 index( HR=1.345, P=0.003), adjuvant therapy( HR=176.652, P=0.012), and operation( HR=0.020, P=0.003), were associated with disease progression. Conclusion:ACC accompanied by hypercortisolism is more prone to invasion and distant metastasis, resulting in shorter survival. Hypercortisolism is an independent prognostic factor for ACC patients.
10.Gastrodin Inhibits Non-alcoholic Fatty Liver Disease via Mediating SREBP1c Signaling Pathway
Menglian ZHANG ; Yaowen ZHANG ; Linfeng TANG ; Zaipeng GONG ; Lan HAN ; Dandan WANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(11):70-77
ObjectiveTo observe the effect of gastrodin on the steroid regulatory element-binding protein 1c (SREBP1c) signaling pathway in high-fat high-cholesterol diet (HFHC)-induced mice and explore the mechanism of gastrodin in the treatment of non-alcoholic fatty liver disease (NAFLD). MethodEight-week-old male C57BL/6J mice were used in vivo and divided into the following four groups, with six mice in each group: normal group, gastrodin group (50 mg·kg-1), model group, and model + gastrodin group (50 mg·kg-1). NAFLD model was established by feeding mice with HFHC for four weeks, and the mice were euthanized and the liver tissues were collected after four weeks. In vitro experiments were performed using Huh7 cells which were divided into five groups, and induced with free fatty acids (FFA, 200 μmol·L-1, oleic acid-palmitic acid 2∶1) to establish an NAFLD cell model. After 24 h, different concentrations of gastrodin (0, 5, 10, 20, and 40 μmol·L-1) were added to each group and cultured for another 24 h. Oil red O staining was used to detect lipid accumulation in mouse liver and Huh7 cells. Hematoxylin-eosin (HE) staining was used to observe pathological changes in liver tissue. Levels of serum total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), and triglycerides (TG) were measured using an automatic biochemical analyzer. Relevant assay kits were used to detect liver TC, TG, and FFA levels. Real-time quantitative polymerase chain reaction (Real-time PCR) and Western blot were used to detect the expression of lipid synthesis-related proteins fatty acid synthase (FASN), acetyl-CoA carboxylase 1 (ACC1), and stearoyl-CoA desaturase 1 (SCD1). ResultCompared with the normal group, the model group showed significantly increased serum TC, LDL-C, and TG levels (P<0.01), liver TC, TG, and FFA levels (P<0.01), increased lipid accumulation in Huh7 cells (P<0.01), and significantly increased expression levels of lipid synthesis-related genes SREBP1c, FASN, ACC1, and SCD1 in mice and Huh7 cells (P<0.01). Compared with the model group, after gastrodin treatment, the serum TC, LDL-C, and TG levels in mice significantly decreased (P<0.05, P<0.01), the severity of fatty liver disease improved significantly, liver TC, TG, and FFA levels decreased significantly (P<0.05, P<0.01), lipid accumulation in Huh7 cells decreased significantly (P<0.05, P<0.01), the expression levels of lipid synthesis-related genes SREBP1c, FASN, ACC1, and SCD1 in mice and Huh7 cells decreased significantly (P<0.05, P<0.01). ConclusionGastrodin can reduce hepatic lipid accumulation and blood lipid levels, improve HFHC-induced NAFLD, and its mechanism of action may be related to the regulation of the SREBP1c lipid synthesis-related signaling pathway.