1.Identification and biological characterization of one Cupriavidus species isolated from human wound
Shiyue KANG ; Chenhui DENG ; Xiaowei CHEN ; Meilian LIN ; Pinghua QU ; Qiang ZHOU
Chinese Journal of Microbiology and Immunology 2024;44(6):510-518
Objective:To analyze the morphology and molecular biology and clarify the taxonomic status of one Cupriavidus species strain SZY C1 isolated from clinical wound specimens. Methods:Strain SZY C1 was subjected to physiological and biochemical identification, 16S rRNA gene sequencing, and whole genome sequencing. Its genomic features and virulence genes were analyzed using bioinformatics software.Results:Strain SZY C1 was a gram-negative, non-fermenting bacterium wihout flagella and the ability to form spores. After culturing on Columbia blood agar plates for 24 h, it formed grayish-white colonies that were round, raised, opaque, and had neatly defined margins. Based on 16S rRNA gene sequence analysis, strain SZY C1 belonged to the genus Cupriavidus with the highest 98.52% similarity to Cupriavidus metallicuns. The genome size of strain SZY C1 was determined to be 5 515 517 bp, with a G+ C content of 67.87%. Whole genome sequencing showed that strain SZY C1 had the closest phylogenetic relationship with Cupriavidus agavae, with an average nucleotide identity value of 84.76% and a digital DNA-DNA hybridization value of 29.1%, which were lower than the identification threshold for prokaryotic species. The strain SZY C1 carried multiple virulence genes, drug resistance genes, and heavy metal resistance genes. Conclusions:Based on phenotypic and genomic analyses, the strain SZY C1 is a potential new species of the Cupriavidus genus.
2.Advances in human skin microecology and microbiome in atopic dermatitis
Ying LIN ; Haimin LUO ; Minling ZHENG ; Wenjun LI ; Pinghua QU
Chinese Journal of Microbiology and Immunology 2020;40(10):800-806
Atopic dermatitis (AD) is a chronic, relapsable and pruritic skin disease, commonly found in children and adolescents. The prevalence of AD is increasing worldwide. It is reported that AD is related to many factors such as genetic inheritance, environment, immunity and skin barrier dysfunction, suggesting a very complex pathogenesis. In recent years, high-throughput technologies in the field of genomics and metabolomics have opened up new perspectives on the pathogenesis of AD, and shown potential application prospects in microbiome transplantation therapies for AD. This review summarized the current advances in the relationship between skin microecology and skin health, the pathogenesis and microbiomic characteristics of AD, features of pathogenic microorganisms, and microbiome transplantation therapies for AD. Based on our own practical experience, we put forward a culturomics research protocol to study the human skin microbiome and a method for quantitative microbiological examination, aiming to provide reference for the prevention, clinical treatment and therapeutic monitoring of AD.
3.Development of a sandwich ELISA for detecting 3AB non-structural protein of foot-and-mouth disease virus.
Yuanfang FU ; Wei HE ; Pu SUN ; Lin YANG ; Huifang BAO ; Yimei CAO ; Xingwen BAI ; Pinghua LI ; Dong LI ; Yingli CHEN ; Lei LIU ; Zengjun LU ; Zaixin LIU
Chinese Journal of Biotechnology 2020;36(11):2357-2366
Antigenic purity is important for quality control of the foot-and-mouth (FMD) whole virus inactivated vaccine. The recommended method for evaluation the antigenic purity of FMD vaccine is to check the serum conversion to non-structural protein (NSP) 3AB antibody after 2 to 3 times inoculation of animals with inactivated vaccine. In this study, we developed a quantitative ELISA to detect the amount of residual 3AB in vaccine antigen, to provide a reference to evaluate the antigenic purity of FMD vaccine. Monoclonal antibody (Mab) of NSP 3A and HRP-conjugated Mab of NSP 3B were used to establish a sandwich ELISA to quantify the NSP 3AB in vaccine antigen of FMD. Purified NSP 3AB expressed in Escherichia coli was serially diluted and detected to draw the standard curve. The detectable limit was determined to be the lowest concentration of standard where the ratio of its OD value to OD blank well was not less than 2.0. Results: The OD value was linearly corelated with the concentration of 3AB protein within the range between 4.7 and 600 ng/mL. The correlation coefficient R² is greater than 0.99, and the lowest detectable limit is 4.7 ng/mL. The amount of 3AB protein in non-purified inactivated virus antigen was detected between 9.3 and 200 ng/mL depending on the 12 different virus strains, whereas the amount of 3AB in purified virus antigen was below the lowest detectable limit. The amount of 3AB in 9 batches of commercial FMD vaccine antigens was between 9.0 and 74 ng/mL, whereas it was below the detectable limit in other 24 batches of commercial vaccine antigens. Conclusion: the sandwich ELISA established in this study is specific and sensitive to detect the content of 3AB protein in vaccine antigen of FMD, which will be a useful method for evaluation of the antigenic purity and quality control of FMD inactivated vaccine.
Animals
;
Antibodies, Viral
;
Enzyme-Linked Immunosorbent Assay
;
Foot-and-Mouth Disease/prevention & control*
;
Foot-and-Mouth Disease Virus
;
Viral Nonstructural Proteins/genetics*
;
Viral Vaccines
4.The Effect of Traditional Culture on the Development of Traditional Chinese Medicine and the Measures of Strengthening the Cultivation of Humanistic Spirit
Pinghua SUN ; Lian LIU ; Zhixiong ZHAO ; Jing LIN ; Jialiang GUO
Chinese Medical Ethics 2017;30(6):697-700
Chinese traditional culture is the soil for the survival and development of traditional Chinese medi-cine,among them,the concept of A thing is valued if it is rare portal factions and master has influenced the development,communication and popularization of traditional Chinese medicine.But the spirit and thinking of harmony unity of heaven and man comparative state benevolence and self-cultivation and self-re-straint has played an active role in the development of traditional Chinese medicine.Thus this paper put forward the suggestions and opinions of strengthening the cultivation of humanistic spirit of higher traditional Chinese medi-cine talents:establishing a comprehensive evaluation standards of higher traditional Chinese medicine talents,es-tablishing scientific development concept to treat the inheritance and innovation of traditional Chinese medicine cul-ture,perfecting the curriculum provision of relevant professions,creating the conducive environment for the trans-mission of traditional Chinese medicine culture,hoping to provide a reference for the explanation of the relationship between traditional culture and traditional Chinese medicine.
5.Relationship between gingival thickness and the underlying bone thickness in maxillary anterior tooth area
Lu LIN ; Pinghua HE ; Sha SU ; Juanjuan ZONG
Journal of Practical Stomatology 2016;32(4):569-572
Objective:To study the relationship between gingival thickness(GT)and the underlying alveolar bone thickness(BT)in maxillary anterior region and the distance from cemento-enamel junction(CEJ)to alveolar crest.Methods:30 young volunteers with healthy gingiva were included.GT was measured at 2mm below the CEJ,buccal BT were measured at 3 locations:2,4 and 6 mm below the alveolar crest respectively,the distance from CEJ to alveolar crest were measured by CBCT and clinical direct measure respectively. Results:The correlation coefficient (r)values between GT and BT at 2,4 and 6 mm below alveolar crest were 0.493,0.383 and 0.342 (P <0.001 )respectively,the r value between GT and the distance from CEJ to alveolar crest was -0.21 3(P <0.01 ).No statistically significant difference was observed between CBCT and clinical measurements(t =-0.521 ,P =0.603).Conclusion:There is positive correlation between GT and BT at 2,4 and 6 mm below alveolar crest and negative relation between GT and the distance from CEJ to alveolar crest.
6.MSCT VRT observation and analysis in the ponticulus of atlas
Pinghua WAN ; Daiping GAO ; Benxing WANG ; Yusheng LIAO ; Runfang LIN
Journal of Practical Radiology 2015;(6):913-916
Objective To expand MSCT obseration and cognition in the ponticulus of atlas,and to improve the diagnosis for it. Methods 263 cases were collected among the patients undergone the examine of MSCT angiography in intracranial and cervical and volume rendering technique(VRT)with unenhance images,and observation and analysis was focused on the posterior arch of atlas. Results 69 cases(97 sides)were detected ponticulus in this group,including unilateral in 41 cases,bilateral in 28 cases.The simple type ponticulus were 82 sides,including the ponticulus posterior (PP)were 60 sides,the ponticulus lateralis (PL)were 1 5 sides,the ponticulus borderland (PB)were 5 sides,The ponticulus middle (PM)were 2 sides.According to the shape of the ponticulus:simple root in 43 sides,opposite beaked in 21 sides,complete type in 18 sides.The ponticulus compound (PC)were 1 5 sides.Conclusion The images of MSCT VRT can showed accurately the ponticulus of atlas,and it can provide reliable imaging evidence on its diagnosis and classification,and it can be used as an unearthly method for its examination.
7.MSCTA observayion of the ditch of vertebral artery of atlas:unnamed osteal structure variation
Pinghua WAN ; Daiping GAO ; Bengxing WANG ; Yusheng LIAO ; Runfang LIN ; Zhiyong LING ; Guiquan CHEN
Journal of Practical Radiology 2015;(10):1605-1607,1623
Objective To explore the belongingness,name and clinical significance of a sort of osteal structure variation (OSV) posterior,outboard and superior to the ditch of vertebral artery of atlas.Methods 23 cases of OSV were collected among 426 patients underwent MSCT intracranial and cervical angiography with volume rendering technique (VRT)to notice the pier point,shape and direction of protuberance,and the relation and influence to the vertebral artery.Results The incidence of OSV was 5.40%(23/426) in which 31 sides were detected,and 1 5 cases in unilateral and 8 cases in bilateral.Simple type was showed at 1 9 sides,compound type at 12 sides with other ponticulus.According to the shape of the ponticulus,simple root was seen at 1 7 sides(13 inferior root and 4 lat-erial root),opposite beaked at 5 sides,complete type at 9 sides.OSV was located at posterior,outbord and superior to the ditch of vertebral artery of atlas and closed to the vertebral artery.The vertebral artery was influenced by OSV as following:in simple OSV vertebral artery stenosis was seen at 4 sides in which unite convulsion was showed at one side;in compound OSV vertebral artery ste-nosis was seen at 5 sides,in which unite convulsion was showed at 2 sides.Conclusion OSV is similar to ponticulus posticus(PP)and ponticulus lateralis(PL),and has similarly importance clinical significance,and should belong to the ponticulus of atlas,and to be named as ponticulus borderland(PB).
8.Identification of Streptobacillus moniliformis isolated from the knee joint pus by 16S rRNA gene sequencing and biochemical reactions
Weizheng ZHANG ; Guangyuan DENG ; Pinghua QU ; Wenke CHEN ; Dongling LIN ; Cha CHEN
The Journal of Practical Medicine 2014;(11):1814-1817
Objective To indentify Streptobacillus moniliformis isolated from the knee joint pus by 16S rRNA gene sequencing and biochemical reactions and explore the clinical value of the method. Methods The bacterial 16S rRNA gene sequence-based identification, bacterial morphology, VITEK 2 automate systems, API 20NE strips, API 20E strips and API 50CH were performed to identify the rare bacteria. Results The bacteria grew slow on blood agar and chocolate agar and were inhibited on Maconkey agar. The bacterial colony on blood agar tookes the form of 1~2 mmomelette, which was translucent and moist with circular protrusion and smooth edges. They were Gram-staining negative and in catenation, its thalli 1~3μm, round, oval or fusiform. Vitek 2 GN-13, API 20NE and API 20E were unable to reach the identification of the bacteria. 16S rRNA gene sequencing showed the bacteria were similar to streptobacillus moniliformis by 100%. Conclusion The rare bacteria isolated from left knee joint are streptobacillus moniliformis. 16S rRNA gene sequences combined with the biochemical reactions is accurate in the identification of these bacteria.
9.Effect of p120 catenin silencing on biological behaviors of PANC-1 cells.
Zhangjun CHENG ; Volker ASSFAG ; Xin SHI ; Shibo LIN ; Jiangyan XIA ; Pinghua YANG ; Norbert HÜSER ; Feng SHEN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2012;32(5):707-712
This study examined the possible role of p120ctn in the pathogenesis and development of pancreatic cancer. PANC-1 cells, a kind of human pancreatic carcinoma cell line, were cultured in this study. p120ctn was immunocytochemically detected in PANC-1 cells. The recombinant lentivirus vector was constructed to knock down the p120ctn expression of PANC-1 cells. Real-time quantitative PCR (RQ-PCR) and Western blotting were used to determine the expression of p120ctn and E-cadherin in PANC-1 cells after p120ctn knockdown. The adhesion, invasion and migration capacity of PANC-1 cells after p120ctn knockdown was detected by cell adhesion, invasion and migration assays. Cell growth was measured by the MTT method. Cell cycle and apoptosis were analyzed by fluorescence-activated cell sorting. The results showed that p120ctn knockdown led to significantly down-regulated E-cadherin and a reduced cell-to-cell adhesion ability in PANC-1 cells. shRNA-mediated knockdown of p120ctn reduced invasion and migration capacity of PANC-1 cells, inhibited cell growth, caused a significant decrease in the percentage of cells in G(1), an increase in S, and promoted apoptosis of PANC-1 cells. It was concluded that p120ctn plays a pivotal role in the proliferation and metastasis of pancreatic carcinoma, suggesting that p120ctn is a novel target for pancreatic carcinoma treatment.
Catenins
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genetics
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Cell Line, Tumor
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Gene Silencing
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Humans
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Pancreatic Neoplasms
;
genetics
10.Effect of p120 catenin silencing on biological behaviors of PANC-1 cells.
Zhangjun, CHENG ; Volker, ASSFAG ; Xin, SHI ; Shibo, LIN ; Jiangyan, XIA ; Pinghua, YANG ; Norbert, HÜSER ; Feng, SHEN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2012;32(5):707-12
This study examined the possible role of p120ctn in the pathogenesis and development of pancreatic cancer. PANC-1 cells, a kind of human pancreatic carcinoma cell line, were cultured in this study. p120ctn was immunocytochemically detected in PANC-1 cells. The recombinant lentivirus vector was constructed to knock down the p120ctn expression of PANC-1 cells. Real-time quantitative PCR (RQ-PCR) and Western blotting were used to determine the expression of p120ctn and E-cadherin in PANC-1 cells after p120ctn knockdown. The adhesion, invasion and migration capacity of PANC-1 cells after p120ctn knockdown was detected by cell adhesion, invasion and migration assays. Cell growth was measured by the MTT method. Cell cycle and apoptosis were analyzed by fluorescence-activated cell sorting. The results showed that p120ctn knockdown led to significantly down-regulated E-cadherin and a reduced cell-to-cell adhesion ability in PANC-1 cells. shRNA-mediated knockdown of p120ctn reduced invasion and migration capacity of PANC-1 cells, inhibited cell growth, caused a significant decrease in the percentage of cells in G(1), an increase in S, and promoted apoptosis of PANC-1 cells. It was concluded that p120ctn plays a pivotal role in the proliferation and metastasis of pancreatic carcinoma, suggesting that p120ctn is a novel target for pancreatic carcinoma treatment.

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