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Objective:To explore the clinical and chest CT features of anti-interferon(IFN)-γ autoantibodies-positive talaromycosis marneffei (TSM) infection in human immune deficiency virus (HIV)-negative patients.Methods:Clinical data and chest CT findings including pulmonary manifestations, bronchial changes, pleural changes, and extrapulmonary manifestations in 54 HIV-negative patients with Talaromyces marneffei (TM) infection and positive anti-interferon-γ(IFN-γ) autoantibody in the First Affiliated Hospital of Guangxi Medical University from December 2012 to September 2019 were retrospectively analyzed. CT score was rated for the degree of lung involvement, and the difference of involvement at different regions of the lung were compared. Results:Of 54 patients, 46 cases had fever, 43 cases had cough and expectoration, 28 cases had cutaneous or subcutaneous lesion, and 19 cases had arthritis or arthralgia. Laboratory tests showed that the value of CD4/CD8 decreased in 29 cases and platelet count increased in 32 cases. Only one of 54 patients had no abnormal findings on chest CT. In the remaining patients, chest CT manifestations were diverse, mainly presenting as fibrous cord-like lesions (87.0%, 47/54), lymph node enlargement (75.9%, 41/54), sporadic nodules (74.1%, 40/54), patchy consolidation (72.2%, 39/54), pleural effusion (59.2%, 32/54), and consolidation lesions associated with air bronchogram (63.0%, 34/54). Most of the lesions showed bilateral distribution (77.8%, 46/54), and involved both peripheral and central regions in 38 cases (70.4%, 38/54). CT score showed that there was no significant difference in the degree of involvement at different regions of the lung.Conclusions:The clinical and imaging manifestations of anti-IFN-γ antibody-positive patients with TM infection are characteristic. Most TSM patients with positive anti-IFN-γ antibody have fever, cough and expectoration. The main features of chest CT are cord focus, nodules, consolidation and pleural effusion. Lymph node enlargement often coexists with the above signs, and most of the lesions are distributed bilaterally.
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Objective: To investigate the anatomical characteristics of the parathyroid lymphatic system and the mechanism of the"negative development"of the carbon nanoparticles for parathyroid gland in thyroidectomy.Methods:This retrospective study used parathyroid tissue samples from patients that were obtained from archival records in the pathology department,including 45 cases of normal parathyroid gland tissues that were accidentally resected in thyroidectomy,10 cases of parathyroid adenomas,and 7 cases of parathyroid carcinoma.Ten cases of normal thyroid tissues were selected as positive control.Immunohistochemistry was performed using the antibodies specific for lymphatic endothelium,such as D2-40 and LYVE-1,and antibodies specific for vascular endothelial cell such as CD31 and CD34,to distinguish them from each other.Results:A total of 62 parathyroid glands samples were stained with vas-cular markers CD31,CD34 and lymphatic markers D2-40,LYVE-1 respectively(partial samples were stained unsuccessfully).Vascular vessels in the CD31 staining group were detected in 50 of 58 examined glands and the positive rate was 86.2%.In the CD34 staining group,positive rate was 100%(60/60).The positive cells were found in the central,periphery and vascular hilum of the glands.Howev-er,lymph vessels in the D2-40 staining group were detected from 17 out of 59 examined glands,with the positive rate of 28.8%;In the LYVE-1 staining group,positive rate was 39.6%(23/58).The positive cells were found in the membrane or vascular hilum,less frequent or undetectable in the central portion.Conclusions:Most of the parathyroid glands of adults might lack a lymphatic network.Only a few adult parathyroid glands had minority lymph vessels,and these lymphatics generally localized at the membrane area or in the vas-cular hilum, which could be one of the main and anatomical mechanisms resulting in drainage failure or obstruction of carbon nanoparticles and thus in parathyroid"negative development."
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Objective To induce skin cancer in BALB/c mice using DMBA as initiator and TPA as tumor promot-er. Through optimizing the doses and frequencies of DMBA administration to establish a stable skin cancer model with less time and causing less skin damage. Methods Shaving the back of mice to expose a piece of skin around 2 cm × 2 cm. The mice were divided into a blank control group and four treatment groups randomly. These four groups were given 1, 2, 4, 7 times 100μg/100μL DMBA/acetone, respectively, in the first week, and twice 4μg/100μL TPA/acetone per week in the next 11 weeks. The body weight changes, time of tumor formation and number of tumors formed were recorded during the experiment. The mice were sacrificed at 12th week and samples of tumor tissue and adjacent normal skin tissue were taken for pathological examination using HE staining. Results Tumors were observed at the 7th week in the group with once DMBA administration in the first week and at the 4th week in the group with twice DMBA administration in the first week. Skin cancers were formed also in the group with 4?time DMBA administration in the first week, however, with signif?icantly more severe skin damages. The mice receiving DMBA everyday in the first week died at the 3th week. Conclusions The best induction protocol for skin cancer in BALB/c mice should be twice DMBA in the first week followed by twice TPA
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AIM: To investigate the relationship between up-regulation of plasminogen activator inhibitor-1 (PAI-1) expression and activation of p38 mitogen-activated protein kinase (p38 MAPK) and extracellular signal-regulated kinase ( ERK) pathways by TGF-β1 in human ovarian cancer cells .METHODS: PAI-1 expression in human ovarian cancer cells treated with TGF-β1 (10 μg/L)was assayed by real-time PCR and Western blotting.The activation of p38 MAPK and ERK was determined by Western blotting using phosphorylated p 38 MAPK and phosphorylated ERK antibodies . Specific p38 MAPK inhibitor (SB203580) or ERK inhibitor (PD98059) was used to inhibit their activation .RESULTS:TGF-β1 up-regulated the expression of PAI-1, and activated p38 MAPK and ERK pathways in the ovarian cancer cells .In-hibition of p38 MAPK activation by SB203580 resulted in significant inhibition of the mRNA expression of PAI-1 induced by TGF-β1.However, inhibition of ERK activation did not significantly alter TGF-β1-induced increase in PAI-1 mRNA level.CONCLUSION: TGF-β1-activated p38 MAPK pathway contributes to the up-regulation of PAI-1 expression by TGF-β1 in ovarian cancer cells .
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OBJECTIVE To investigate the antidepressant effect of triptolide in chronically unpredictable stressed mice and its possible protective effect on brain derived neurotrophic factor(BDNF). METHODS One method was selected from 8 different stress methods each day,and the mice were treated with triptolide(20,40,80 and 160μg·kg-1)10 min before the stress method. A chroni?cally unpredictable stressed model was established and after 14 d stress experiment, the total distance in the locomotor activity and the immobility time in the force swimming test and tail test were observed respectively. Triptolide(20,40,80 and 160μg·kg-1)was given 10 min before the test. In addition, Western blot was used to analyze the expression of phosphorylated cAMP response element binding protein(p-CREB) and BDNF in the hippocampus and frontal cortex. RESULTS There was no effect on the locomotor activity in any group. Compared with the normal control group,the chronically unpre?dictable stressed group showed obvious depressive-like behavior,while the immobility time in the force swimming test decreased from(161 ± 18)s in chronically stressed group to(102 ± 14)s(P<0.05) and(83±14)s(P<0.01)when mice were ip given triptolide 80 and 160μg·kg-1, respectively,and(77± 11)s(P<0.01)in imipramine(IMI)hydrochloride group(10 mg?kg-1),and(96±9)s(P<0.01)in fluox?etine(FLU)group(10 mg?kg-1). The immobility time in the tail suspension test decreased from(128± 8)s in chronically stressed group to(93±9)s(P<0.05),(85±8)s(P<0.01)and(77±11)s(P<0.01)when mice were ip given triptolide 40,80 and 160μg · kg-1 respectively,and(64 ± 9)s(P<0.01)in IMI hydro?chloride 10 mg?kg-1 group,and(72±6)s(P<0.01)in FLU group(10 mg?kg-1). Moreover,the expression of p-CREB in the hippocampus and frontal cortex significantly increased in triptolide 80 and 160μg·kg-1 groups(P<0.05),so did the expression of BDNF in the hippocampus and frontal cortex in triptolide 80 and 160μg·kg-1 groups(P<0.05). CONCLUSION Triptolide can ameliorate the depressive-like behavior in chronically unpredictable stressed mice,which may be related to the cAMP-CREB-BDNF signal transduction cascades.
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Objective To construct short hairpin RNA (shRNA) recombinant expression vector for herpes simplex virus typeⅡ(HSV-2) UL29 gene and observe its inhibitory effect on HSV-2. Methods Four interference target sites of HSV-2UL29 gene were selected to construct 4 groups of small hairpin RNA respectively,named shRNA recombinant expression vector. The expression vectors were transfected into HEK293 cells with liposome. HEK293 cells were infected with HSV-2 after expression vector being transfected. The viral titer was estimated by end-point titration assay. The level of transcription was estimated by Real-Time PCR method. The expressing effect of protein was detected by Western-blot. Results Recombinant expression vector pGPU6/GFP/Neo-shRNA was constructed successfully. The result of end-point titration assay showed that the viral titer was reduced comparing with blank control (P<0.05). The result of RT-PCR showed that inhibition rates were respectively 28.80%, 59.95%, 66.08%and 36.27% comparing with blank control, and there were significant differences (P < 0.05). The effect of UL29shRNA1461 group was the best one. The result of Western-blot showed that the expressing quantity of ICP8 was reduced. Conclusion Recombinant expression vector pGPU6/GFP/Neo-shRNA can interfere HSV-2 UL29 gene expression from different cell level in vitro, which can inhibit the replication of HSV-2 genome in HEK293 cells. Thus, RNA interference (RNAi) is conducive to the further exploration of viral therapy.
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Objective To clarify the role of claudin-4 in endometrial tumorigenesis and explore claudin-4 be as potentially useful agent in the treatment of endometrial carcinoma.Methods The expression of claudin-4 in 62 endonetrioid endometrial carcinoma (EEC),30 atypical hyperplasia endometrial tissue and 60 human normal endometrium was determined using immunohistochemistry and realtime PCR.Ninety female BALB/c mice were transplanted with Ishikawa endometrial cancer cells,which were divided into three groups with different intraperitoneal treatments with cisplatin,paclitaxel and saline solution.After the observation period,the tumors were extracted and stained with monoclonal antibody against claudin-4.The messenger RNA expression of claudin-4 was also detected using real-time PCR.Results Among the EEC samples,34％ (21/62) showed medium staining for claudin-4 and 66％ (41/62) showed intense staining.In atypical hyperplasia group,27％ (8/30) showed weak staining,53％ (16/30) showed medium staining and 20％ (6/30) showed intense staining for claudin-4.Of the normal endometrial tissue,47％ (28/60) showed weak staining and 53％ (32/60) showed no staining for claudin-4.According to real-time PCR,the relative quantity of claudin-4 was 170 ± 12 in EEC group,89 ± 15 in atypical hyperplasia group and 18 ± 3 in normal endometrium.Compared with those in atypical hyperplasia group and normal endometrium group,the protein and mRNA expression of claudin-4 were significantly increased in the group of EEC (all P ＜ 0.05).In the study of Ishikawa xenografts,no significant changes in tumor volume and claudin-4 expression were shown in paclitaxel group compared with that in the control group.Nevertheless,a significant reduction of the tumor growth and a significant decrease in claudin-4 expression were observed in cisplatin group.After cisplatin treatment,the tumor volume was significantly decreased [(0.51 ±0.21) versus (0.73 ±0.12) cm3],and the mRNA expression of claudin-4 was also significantly decreased (153 ± 35 versus 273 ± 27).Conclusion These results demonstrate that claudin-4 is strongly expressed in EEC,which may be a useful biomarker to monitor the effects of chemotherapy in patients with endometrial carcinoma.
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ObjectiveTo explore the regulation of claudin-4 expression in endometrial adenocarcinoma cell lines by progesterone.Methods Ishikawa cells were treated with various concentrations of megestrol acetate (MA:2,5,10,15,20 mg/L).After cultured for 24,48 and 72 hours,cells growth were measured by methyl thiazolyl tetrazolium (MTT).The group of Ishikawa cells incubated with MA at the 50％ inhibitory concentration ( IC50 ) was selected for cell apoptosis assay by using transmission electron microscopy and flow cytometry method.Real-time PCR and western blot were used for detecting the mRNA and protein expression levels of claudin-4.The localization of claudin-4 was examined by immunofluorescent staining.Results The inhibitory effects of megestrol acetate on the growth of Ishikawa cells were dosedependent and time-dependent.IC50 of MA on Ishikawa cells was 15 mg/L after incubated for 72 hours.After MA treatment,Ishikawa cells showed shrinkage,nuclear chromatin condensation,fractures of nuclear membrane and endoplasmic reticulum expansion,even round apoptotic bodies were found.The apoptosis rate of cells before MA treatment was (0.076 ±0.024)％,and the rate was (3.934 ±0.816)％ by MA treated for 72 hours,in which there were signicant difference( P ＜ 0.05 ).The relative quantification of claudin-4 mRNA and protein of the cells before MA treatment were 0.64 ± 0.20 and 0.94 ± 0.18,while they were 0.47 -0.15 and 0.62 ±0.15 after MA treated.The expression of claudin-4 was significantly decreased after MA treatment ( P ＜ 0.05 ).The localization of claudin-4 transferred from cytomembrane to cytoplasm and nucleus after MA treatment.Conclusions MA could inhibite the growth of Ishikawa cells,in which the mechanism may be decrease the expression of claudin-4 and the apoptosis of cells.The distribution change of claudin-4 may be related to the anti-cancer effect of progesterone.
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Objeetive To investigate the expression of claudin-4 in the eutopic and ectopic endometfium of women with endometriosis and evaluate the role of clandin-4 in the pathogenesis of endometriosis.Methods Thiny-five women with endometriosis and 35 controls were studied.Expression of elaudin-4 was investigated using immunohistochemistry,western blot and RT.PCR,respectively.Morphologic change of tight junction was also observed in different kinds of endometria.Results (1)G1andular epithelial cells of control endometrium and eutopic endometrium showed intact tight iunctions in electron micrographs,whereas the morphology of tight junctions in ovarian endometriotic tissue was disrupted and collagen bundles could be easily detected.(2)The immunohistochemical staining of claudin-4 was localized to the glandular epithelial cell membrane.Deftcient or weak staining Wag found in ovarian endometriotic tissues. In control endometrium.eutopic and ectopic endometrium of women with endometriosis,the expression of clandin-4 protein Was 89±24.84±22 and 27±14.respectively.Relative expression of claudin-4 mRNA Was 14.5±6.8,13.8±9.5 and 2.6±2.5.respectively.Expression of claudin-4 Was significantly lower in the ectopic endometriotic tissue than in the eutopic cndometrium and the control at both mRNA and protein levels(P<0.05). N0 significant difference Was foand between eutopic endometrium from women with endometriosis and control endometrium from women without endometriosis (P>0.05).Conclusion Down-regulated expression of claudin-4 might play a pathogenic role in the formation of endometriosis.
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AIM:To investigate the expressions of methylthioadenosine phosphorylase(MTAP)and ornithine decarboxylase(ODC)in human ovarian cancer.METHODS:60 fresh samples of ovarian cancer were collected.The expressions of MTAP mRNA and protein were analyzed by using RT-PCR and Western blotting,respectively.ODC activity was measured by high performance liquid chromatography.RESULTS:The expression levels of MTAP mRNA and protein in ovarian cancer were lower than those of control.In 9 of the 60 samples(15%)there were absence of detectable MTAP mRNA and protein.No significant relevance was found between the expression of MTAP and clinical pathologic features.ODC activity in ovarian cancer was(3.82?1.03)U,which was higher than that of normal ovarian tissues(1.38?0.59)U.ODC activity was related with tumor grade.In MTAP-deficiency ovarian cancer tissues ODC activity was significantly increased when compared with that of MTAP-expressing ovarian cancer samples.CONCLUSION:Down-regulated MTAP expression and up-regulated ODC activity really exist in ovarian cancer.Activation of ODC resulting from MTAP deletion may be one of the pathogenetic factors of ovarian cancer.