1.Differential analysis of intestinal flora in patients with hepatic blastomycosis based on second-generation sequencing
Xiaoya MA ; Xiangren A ; Jideng MA ; Jianwu ZHOU ; Peng CHENG ; Yun TANG
Chinese Journal of Preventive Medicine 2025;59(1):101-109
Exploring the variability of the intestinal flora of patients with hepatic blastocysticercosis and searching for members of the intestinal microflora that may play a role in the disease process by means of macro-genome sequencing technology. A case-control study was used to include fecal samples from patients with hepatic vesicular schistosomiasis admitted to Qinghai Provincial People′s Hospital between October 2023 and January 2024 and individuals attending health checkups. The experimental group (AE group) consisted of 10 patients with liver vesicular schistosomiasis and the control group (NC group) consisted of 9 individuals attending health checkups. Macrogenomic sequencing was performed on these two groups of samples using the Illumina Novaseq 6000 sequencing platform, using fastp (v0.20.1) to remove junctions, and bbmap (v38.93-0) to remove the hosted sequences, followed by sequence splicing using MEGAHIT (v1.2.9), and then using prodigal (v2.6.3) to The spliced scaffold was subjected to ORF prediction and translated into amino acid sequences, followed by the construction of a non-redundant gene set using MMSeqs2 (v13.45111), and finally compared with the non-redundant gene set using salmon (v1.8.0). Species were annotated by the non-redundant database, species abundance was calculated in each sample, and the two sets were tested using Wilcoxon rank sum test. Finally, the differences in intestinal flora between the two groups were statistically analyzed using linear discriminant analysis, and the correlation between the differential intestinal flora and clinical indicators was analyzed using redundancy analysis (RDA). The results showed that the effective data volume of each sample was distributed from 10.41 to 12.46 G. The number of ORFs in the de-redundantly constructed gene catalogue (non-redundant gene set) was 4 951 408, and the annotation rate of the non-redundant genes was 97.97% when compared with the NR database. The ages of the study subjects in the two groups were (44.78±4.58) years in the NC group and (42.90±10.44) years in the AE group, and the difference was not statistically significant ( t=0.530, P=0.476). The two groups were matched for body mass index (BMI) ( t=2.368, P=0.142), gender ( χ2=0.200, P=0.655), and dietary habits. There was no statistically significant difference in alpha diversity in the AE group (ACE index, t=0.942; chao1 index, t=0.947; shannon index, t=0.813, the simpson′s index, t=0.613, P>0.05), while beta diversity analysis showed significant differences in the overall structure of the two communities (Stress=0.054 5). A total of 120 species were annotated at the phylum level, of which two differed. While 1 736 species were annotated at the genus level, 69 were different, and 309 were different at the species level. The AE group ranked the top 6 in terms of abundance of Anaplasma, Escherichiaceae, Clostridium, Alternaria, Ruminalia, and Treponema spp. at the genus level; whereas, Segatella, Prevotella, E. faecalis, Rossella, and beneficial rod-shaped bacteria were more abundant in the NC group. There were differences in the abundance and diversity of intestinal flora between the two groups, and the structure of community composition was significantly different. Statistical results by linear discriminant analysis (LDA) showed that LDA scores >2 in the NC group included beneficial bacillus spp. and E. faecalis spp. in young infants, etc. LDA scores >2 in the AE group at the mid-species level included Clostridium polterococcus, unknown microorganisms in the genus Clostridium intestinalis, Hathaway′s Henkett′s bacillus, and Clostridium oryzae in the genus Clostridium refractory to culture and small Clostridium spp. in the AE group. Clostridium intestinalis. The RDA results showed a negative correlation between beneficial rod genera and liver function indices, and a positive correlation between Clostridium intestinalis genera and liver function indices. In conclusion, patients with hepatic blastomycosis have altered intestinal flora abundance and diversity, with significant structural changes in community composition and differences in several genera, including Mycobacterium anisopliae and Clostridium intestinalis, and imbalances in the intestinal flora may affect hepatic function by influencing intestinal metabolites and may have an impact on the development of hepatic blastomycosis, a finding that warrants further in-depth study.
2.The effect of ethyl alcohol extract of moutan cortex on virulence factors of Candida tropicalis
Yun TANG ; Xiangren A ; Peng CHENG ; Jianwu ZHOU ; Jideng MA ; Xiaoya MA
Chinese Journal of Laboratory Medicine 2025;48(2):258-264
Objective:To investigate the in vitro bacteriostatic effect of ethyl alcohol extract of Moutan Cortex (EAEMC) on Candida tropicalis and its effect on virulence factors, including aspartic protease, hemolysin, phospholipase, esterase, lipase activities and biofilm. Methods:EAEMC powder was obtained by ultrasonic extraction, decompression concentration and lyophilization; the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of EAEMC on 21 clinical strains and one standard strain of Candida tropicalis were determined by microdilution. Five extracellular enzyme activities of Candida tropicalis and the effect of EAEMC on them were detected by the plate assay, and the results were analyzed by ANOVA. The biofilm model of Candida tropicalis was constructed in vitro, and the inhibition rate of EAEMC on Candida tropicalis biofilm was evaluated using the thiazolyl blue (MTT) method. Results:The MIC of EAEMC against Candida tropicalis BNCC335988 was 12.5 g/L and the MBC value was 25 g/L, while for the clinical strains, the MIC was 12.5-25 g/L and the MBC was 25-50 g/L. Aspartic protease, esterase and hemolytic activities of Candida tropicalis were positive, but phospholipase and lipase showed negative activities. At a concentration of 1/2 MIC of EAEMC, the aspartic protease and hemolytic activities of Candida tropicalis were completely inhibited the aspartic protease and hemolytic activities of Candida tropicalis were completely inhibited and the esterase activity was completely inhibited at a concentration of MIC of EAEMC. The inhibition of Candida tropicalis BNCC335988 biofilm by EAEMC reached more than 70% at a concentration of 2MIC, more than 80% at a concentration of 4MIC, and more than 90% at a concentration of 8MIC. Conclusion:EAEMC can achieve bacteriostatic effects by reducing the aspartic protease, esterase and hemolysin activities of Candida tropicalis, as well as inhibiting biofilm formation.
3.Differential analysis of intestinal flora in patients with hepatic blastomycosis based on second-generation sequencing
Xiaoya MA ; Xiangren A ; Jideng MA ; Jianwu ZHOU ; Peng CHENG ; Yun TANG
Chinese Journal of Preventive Medicine 2025;59(1):101-109
Exploring the variability of the intestinal flora of patients with hepatic blastocysticercosis and searching for members of the intestinal microflora that may play a role in the disease process by means of macro-genome sequencing technology. A case-control study was used to include fecal samples from patients with hepatic vesicular schistosomiasis admitted to Qinghai Provincial People′s Hospital between October 2023 and January 2024 and individuals attending health checkups. The experimental group (AE group) consisted of 10 patients with liver vesicular schistosomiasis and the control group (NC group) consisted of 9 individuals attending health checkups. Macrogenomic sequencing was performed on these two groups of samples using the Illumina Novaseq 6000 sequencing platform, using fastp (v0.20.1) to remove junctions, and bbmap (v38.93-0) to remove the hosted sequences, followed by sequence splicing using MEGAHIT (v1.2.9), and then using prodigal (v2.6.3) to The spliced scaffold was subjected to ORF prediction and translated into amino acid sequences, followed by the construction of a non-redundant gene set using MMSeqs2 (v13.45111), and finally compared with the non-redundant gene set using salmon (v1.8.0). Species were annotated by the non-redundant database, species abundance was calculated in each sample, and the two sets were tested using Wilcoxon rank sum test. Finally, the differences in intestinal flora between the two groups were statistically analyzed using linear discriminant analysis, and the correlation between the differential intestinal flora and clinical indicators was analyzed using redundancy analysis (RDA). The results showed that the effective data volume of each sample was distributed from 10.41 to 12.46 G. The number of ORFs in the de-redundantly constructed gene catalogue (non-redundant gene set) was 4 951 408, and the annotation rate of the non-redundant genes was 97.97% when compared with the NR database. The ages of the study subjects in the two groups were (44.78±4.58) years in the NC group and (42.90±10.44) years in the AE group, and the difference was not statistically significant ( t=0.530, P=0.476). The two groups were matched for body mass index (BMI) ( t=2.368, P=0.142), gender ( χ2=0.200, P=0.655), and dietary habits. There was no statistically significant difference in alpha diversity in the AE group (ACE index, t=0.942; chao1 index, t=0.947; shannon index, t=0.813, the simpson′s index, t=0.613, P>0.05), while beta diversity analysis showed significant differences in the overall structure of the two communities (Stress=0.054 5). A total of 120 species were annotated at the phylum level, of which two differed. While 1 736 species were annotated at the genus level, 69 were different, and 309 were different at the species level. The AE group ranked the top 6 in terms of abundance of Anaplasma, Escherichiaceae, Clostridium, Alternaria, Ruminalia, and Treponema spp. at the genus level; whereas, Segatella, Prevotella, E. faecalis, Rossella, and beneficial rod-shaped bacteria were more abundant in the NC group. There were differences in the abundance and diversity of intestinal flora between the two groups, and the structure of community composition was significantly different. Statistical results by linear discriminant analysis (LDA) showed that LDA scores >2 in the NC group included beneficial bacillus spp. and E. faecalis spp. in young infants, etc. LDA scores >2 in the AE group at the mid-species level included Clostridium polterococcus, unknown microorganisms in the genus Clostridium intestinalis, Hathaway′s Henkett′s bacillus, and Clostridium oryzae in the genus Clostridium refractory to culture and small Clostridium spp. in the AE group. Clostridium intestinalis. The RDA results showed a negative correlation between beneficial rod genera and liver function indices, and a positive correlation between Clostridium intestinalis genera and liver function indices. In conclusion, patients with hepatic blastomycosis have altered intestinal flora abundance and diversity, with significant structural changes in community composition and differences in several genera, including Mycobacterium anisopliae and Clostridium intestinalis, and imbalances in the intestinal flora may affect hepatic function by influencing intestinal metabolites and may have an impact on the development of hepatic blastomycosis, a finding that warrants further in-depth study.
4.The effect of ethyl alcohol extract of moutan cortex on virulence factors of Candida tropicalis
Yun TANG ; Xiangren A ; Peng CHENG ; Jianwu ZHOU ; Jideng MA ; Xiaoya MA
Chinese Journal of Laboratory Medicine 2025;48(2):258-264
Objective:To investigate the in vitro bacteriostatic effect of ethyl alcohol extract of Moutan Cortex (EAEMC) on Candida tropicalis and its effect on virulence factors, including aspartic protease, hemolysin, phospholipase, esterase, lipase activities and biofilm. Methods:EAEMC powder was obtained by ultrasonic extraction, decompression concentration and lyophilization; the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of EAEMC on 21 clinical strains and one standard strain of Candida tropicalis were determined by microdilution. Five extracellular enzyme activities of Candida tropicalis and the effect of EAEMC on them were detected by the plate assay, and the results were analyzed by ANOVA. The biofilm model of Candida tropicalis was constructed in vitro, and the inhibition rate of EAEMC on Candida tropicalis biofilm was evaluated using the thiazolyl blue (MTT) method. Results:The MIC of EAEMC against Candida tropicalis BNCC335988 was 12.5 g/L and the MBC value was 25 g/L, while for the clinical strains, the MIC was 12.5-25 g/L and the MBC was 25-50 g/L. Aspartic protease, esterase and hemolytic activities of Candida tropicalis were positive, but phospholipase and lipase showed negative activities. At a concentration of 1/2 MIC of EAEMC, the aspartic protease and hemolytic activities of Candida tropicalis were completely inhibited the aspartic protease and hemolytic activities of Candida tropicalis were completely inhibited and the esterase activity was completely inhibited at a concentration of MIC of EAEMC. The inhibition of Candida tropicalis BNCC335988 biofilm by EAEMC reached more than 70% at a concentration of 2MIC, more than 80% at a concentration of 4MIC, and more than 90% at a concentration of 8MIC. Conclusion:EAEMC can achieve bacteriostatic effects by reducing the aspartic protease, esterase and hemolysin activities of Candida tropicalis, as well as inhibiting biofilm formation.
5. The level of formaldehyde in the air of the anatomic laboratories and its prevention measures
Hongmei QIU ; Jianwu CHEN ; Li TANG ; Hui WANG ; Wanyu DU ; Lin YANG ; Yuxing DU
China Occupational Medicine 2018;45(06):723-726
OBJECTIVE: To investigate the level of formaldehyde in the air of anatomical laboratories and to explore the effect of different prevention measures on reducing the level of formaldehyde. METHODS: The level of formaldehyde in two morphological laboratories of a medical college of a university were measured with a comprehensive atmospheric sampler at different physical ventilation time,different time after spraying biological enzymes and different masks. RESULTS: The level of formaldehyde decreased with the increase of ventilation time in front,middle and back of the laboratory( P < 0. 05).The level of formaldehyde at 30 and 60 minutes after spraying bio-enzymes in three parts of the laboratory was lower than that at 0 minute in the same part of the laboratory( P < 0. 05),and lower than that in the bio-enzymes non-spraying laboratory at the same part and same time point( P < 0. 05). At 0,30 and 60 minutes ventilation time points,the level of formaldehyde in the activated carbon mask group was lower than that in the normal sampling group,the non-woven mask group and the defatted gauze mask group at the same time point( P < 0. 05). CONCLUSION: Both physical ventilation and bio-enzyme degradation can reduce indoor formaldehyde pollution. Active carbon mask can achieve good protection effect.
6.Prepartation of Gelsolin-targeted ultrasound contrast agent and experiment in vitro
Haocheng QIN ; Jun WU ; Meng ZHOU ; Yuhong ZHANG ; Yu SONG ; Jieming LI ; Xiaona WEN ; Jianwu TANG ; Haitao RAN
Chinese Journal of Medical Imaging Technology 2017;33(6):826-831
Objective To prepare a kind of Gelsolin-targeted ultrasound contrast agent (GSN-PLGA) and to explore its targeting and imaging effection in vitro.Methods The high molecular PLGA-COOH ultrasound contrast agents were prepared by a modified double emulsion technique and then conjugated with Gelsolin monoclonal antibody by carbodiimide technique.The physical property of contrast agent was determined.And the connectivity condition of ultrasound contrast agent with Gelsolin monoclonal antibody was estimated.The targeting ability and the effect of enhancing ultrasound imaging in vitro were explored.Results The average diameter of GSN-PLGA was (575.67 ± 4.71) nm.The potential was (-11.46±1.19) mV.And the binding rate of Gelsolin monoclonal antibody was 96.93%.In vitro experimentshowed more GSN-PLGA could be intaked by Hca-F cells and the ultrasound imaging cloud be enhanced greatly.Conclusion The GSN-PLGA nanoparticle can bind to Hca-F cells specifically and can enhance the ultrasound imaging greatly.
7.Expression levels of Ech1 in hepatocarcinoma and subcellular location
Jun ZHANG ; Hong QU ; Jun MAO ; Yuanyi WEI ; Yuhong HUANG ; Huaxin WANG ; Jianwu TANG
Chinese Journal of Clinical and Experimental Pathology 2015;(3):247-250
Purpose To study the expression of Ech1 in hepatocarcinoma and its clinical significance, and to explore the relationship between subcellular location of Ech1 and malignant biologic behaviour of hepatocarcinoma. Methods Immunohistochemical analysis was used to detect Ech1 expression in the 20 cases of normal liver tissue and the 30 cases of hepatocarcinoma. Subcellular location of Ech1 in Hca-F and FEch1-down cell was observed with subcellular protein extraction. Results The expression of Ech1 in primary hepa-tocarcinoma was increased compared to that of normal liver tissues ( P<0. 05 ) , while there was no significant difference between the expression of Ech1 with the age, AFP, and with or without liver cirrhosis or hepatitis virus in hepatocarcinoma (P>0. 05). Ech1 was found expressed almost at the same location although the expression level one is normal and the other is downregulation. Ech1 expres-sion was found in cytosol, membrane fraction, and its expression was higher in cytosol than other fractions both in Hca-F cell and Ech1 downregulated Hca-F cell. Conclusions The expression of Ech1 in primary hepatocarcinoma was increased, which may indicate that Ech1 is a critical factor in the development of primary hepatocarcinoma, but the subcellular location of Ech1 has not much contribution to that.
8.The expression and signiifcance of ERp29 in 2 mice hepatocellular carcinoma ascites syngeneic cell lines with high and low lymph node metastasis rates
China Oncology 2015;(2):95-98
Background and purpose:ERp29 belongs to endoplasmic reticulum stress proteins and might play roles in neoplasm metastasis. This study aimed to investigate the expression of ERp29 in Hca-F and Hca-P cells and to elucidate its role in lymphatic metastasis.Methods:Immunohistochemistry, Western blot and lfow cytometry analysis were used to detect the expression of ERp29 in Hca-F and Hca-P.Results:The results of immunohistochemistry suggested that ERp29 protein was located at cytoplasm of hepatic cells and some were also detected in the nucleus. The results of western blot suggested that ERp29 had positive expression in Hca-F and Hca-P cells. Its expression in Hca-F cells was apparently lower than that in Hca-P cells. And there was statically different between Hca-F and Hca-P cells (P<0.01). The relative lfuorescence intensity of ERp29 protein was signiifcantly lower in Hca-F cells (375.27±47.33) than that in Hca-F cells (623.91±46.80) by lfow cytometry (P<0.01).Conclusion:The different expression of ERp29 may be related to the potential ability of tumor lymphatic metastasis in Hca-F and Hca-P cells.
9.Apoptosis of MDA-MB-231 cells induced by berberine alpha-hydroxy beta-decanoylethyl sulfonate.
Acta Pharmaceutica Sinica 2014;49(1):131-5
To investigate the cell proliferation inhibition and apoptosis induced by berberine a-hydroxy f-decanoylethyl sulfonate (HB) on MDA-MB-231 cells in vitro, and the inhibitory effect of HB on the expression of poly adenosine diphosphate RNA polymerase (PARP), MTT assay was used to detect the viability of MDA-MB-231 cells and cell cycle was examined by flow cytometry. The results showed that HB could significantly inhibit the proliferation of MDA-MB-231 cells, and mildly arrested cell cycle progression at S phase. The IC50S for 24, 48 and 72 h treatment were 4.65, 1.46 and 0.75 mg.L-1 (7.55, 2.37 and 1.22 micromol.L-1), respectively. Annexin V-FITC/PI double staining assay showed that HB increased apoptotic ratio of MDA-MB-231 cells. Western blotting analysis showed the expressions of procaspase-3, procaspase-9 and PARP were decreased after HB treatment, while their fragment increased. The results suggest that HB can inhibit the growth and induce apoptosis of MDA-MB-231 cells, which may be associated with inhibition of the expression of procaspase-3, procaspase-9 and PARP.
10.Relationship between activated STAT3 protein and epithelial-mesenchymal transition in papillary thyroid carcinoma.
Zhuqing ZHANG ; Yan BAI ; Ping LI ; Junjun ZHAO ; Yanxia WANG ; Lei SUN ; Jianwu TANG
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2013;27(22):1265-1268
OBJECTIVE:
To investigate the expression of signal transducer and activator of transcription 3 (STAT3) and phosphorylated STAT3 (p-STAT3) protein in papillary thyroid carcinoma (PTC), and to explore the correlation and significance between the expression of STAT3, p-STAT3 and epithelial-mesenchymal transition (EMT).
METHOD:
The expression of STATS3. p-STAT3, E-cadherin and Vimentin protein in 56 cases of PTC specimens and adjacent normal tissues specimens ware detected by immunohistochemistry. The correlation of the expression of STATS, p-STAT3, E-cadherin and Vimentin protein in PTC with clinicopathological characteristics was analyzed.
RESULT:
The positive rates of STAT3, p-STAT3 in PTC tissue were significantly higher than those in adjacent normal tissues specimens respectively (P < 0.01). The positive rates of E-cadherin in PTC tissues were remarkably lower, compared to adjacent normal tissues specimens (P < 0.01), however the positive rates of Vimentin in PTC tissues were remarkably higher, compared to adjacent normal tissues specimens (P < 0.01). The expression of STAT3, p-STAT3, E-cadherin and Vimentin protein were associated with lymph node metastasis and clinical stage (all P < 0.05). The expression of STAT3 and p-STAT3 was negatively correlated with E-cadherin expression (r = -0.494 and r = -0.364, P < 0.01, respectively), but positively with Vimentin expression (r = 0.533 and r = 0.377, P < 0.01, respectively) in PTC tissues.
CONCLUSION
PTC tissues have STAT3 protein activation and EMT phenotype, as were all correlated significantly with PTC invasion and metastasis. STAT3 signaling pathway activation might mediate EMT and then promote PTC invasion and metastasis.
Adult
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Antigens, CD
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Cadherins
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metabolism
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Carcinoma
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metabolism
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pathology
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Carcinoma, Papillary
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Epithelial-Mesenchymal Transition
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Female
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Humans
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Male
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Middle Aged
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Phosphorylation
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STAT3 Transcription Factor
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metabolism
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Thyroid Cancer, Papillary
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Thyroid Neoplasms
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metabolism
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pathology
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Vimentin
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metabolism

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