1.Analysis of the key amino acids involved in the function and cellular self-assembly of FtsZ protein in Escherichia coli strains
Xiaowei ZHENG ; Qiaonan LU ; Yujia HUO ; Yuanfang MA ; Feng LU
Chinese Journal of Microbiology and Immunology 2015;(4):241-246
Objective To investigate the self-assembly and cellular localization patterns of fila-mentous temperature-sensitive protein Z (FtsZ) in Escherichia coli (E.coli) strains by using FtsZP74R, FtsZG77D and FtsZA81R mutants.Methods YFP or His labeled FtsZ proteins and the plasmids of FtsZ mu-tants were constructed by using molecular clone and site-directed mutagenesis methods.The targeted proteins were purified by affinity chromatography.FL37(△ftsZ-Cat) strains were constructed via linear DNA homol-ogous recombination.Living cell imaging was performed to observe the cellular localization patterns of FtsZ protein and its mutants in E.coli strains.The interactions between FtsZ-FtsZ/FtsZ mutants were examined by coi-mmunoprecipitation assay . The polymerization properties of FtsZ mutants were analyzed by light scattering.The activities of GTPase were monitored by using high performance liquid chromatography.Re-sults The P74, G77 and A81 amino acids were respectively replaced by different polar amino acids to change the amphipathicity of the helix within the domain of FtsZ ( 74-82 ) .The YFP-labeled FtsZP74R , FtsZG77D and FtsZA81R mutants failed to assemble into functional Z-ring structure in E.coli strains.The inter-actions between FtsZ protein and its mutants were weakened or completely disappeared.In addition, in vitro experiments showed that P74R, G77D and A81R mutations caused a decrease in the polymerization efficien-cy of FtsZ monomer.The activity of GTPase was significantly decreased in the FtsZA81R mutant. Conclusion The P74, G77 and A81 were critical amino acids in the function and assembly of FtsZ protein in E.coli strains.Moreover, A81 amino acid regulated the lateral interaction of FtsZ monomer and the activity of GTPase.
2.A pilot molecular mechanism study on lacrimal gland as a potential human immunodeficiency virus reservoir
Ziyang, LIU ; Xiaowei, LIU ; Junjie, YE ; Yang, HAN ; Zhen, HUO
Chinese Journal of Experimental Ophthalmology 2017;35(5):410-415
Background Acquired immune deficiency syndrome (AIDS) is an infectious disease caused by human immunodeficiency virus (HIV).Highly active antiretroviral therapy (HAART) is an effective treatment for AIDS,but it cannot completely eliminate the viral load in the body for the existence of HIV reservoir.Previous studies demonstrated that HIV could be detected in tears of virus load negative AIDS patients who received effective HAART,suggesting that lacrimal gland is another member of HIV reservoirs.Objective The aim of this study was to explore whether lacrimal gland has a molecular basis of HIV infection and the mechanism of lacrimal gland infection of HIV.Methods Fourteen specimens of lacrimal gland were collected during the surgery from 14 patients with lacrimal gland diseases in Peking Union Medical College Hospital from November 2013 to December 2015,including 13 non-HIV-infected patients and 1 HIV-infected patient.In 13 non-HIV infected patients,lacrimal glands prolapse was in 12 patients with the normal pathological tissue structure and dacryoadenitis was in 1 patient with the histopathological diagnosis of interstitial lymphoid tissue hyperplasia.The clinical manifestation of HIV-infected patient was dacryoadenitis with the histopathological diagnosis of interstitial lymphoid tissue hyperplasia.The paraffin sections of 12 non-HIV-infected specimens and 1 HIV-infected specimen were prepared,and the expressions of CD4,C-X-C chemokine receptor 4 (CXCR4) and C-C chemokine receptor type 5 (CCR5) in lacrimal gland specimens were detected by immunohistochemistry and verified in 1 specimen of non-HIV-infected specimen by immunofluorescence technology.Results Immunohistochemistry showed that CD4 was suspiciously positive expression in non-HIV-infected specimens with the strong background staining.CXCR4 was positively expressed in cytoplasm and nuclei of most lacrimal epithelial cells of lacrimal gland epithelial cells in each specimen,and CCR5 was focally expressed in few lacrimal gland epithelial cells in each specimen.In addition,CD4,CXCR4 and CCR5 were positively expressed in intercellular scattered lymphocytes on the specimens.Immunofluorescence assay showed that CD4,CXCR4 and CCR5 were expressed in the specimens with the red fluorescence,with the linear-and patchy-like distribution mainly in cellular membrane for CD4 or spot-like distribution for CXCR4 and CCR5 in the cytoplasm.Conclusions HIV receptor CD4 and accessory receptor CXCR4,CCR5 are positively expressed in the lacrimal gland epithelial cells,which is the molecular basis of HIV infection and become a potential HIV reservoir preventing HIV eradication.
3.Effects of triptonoterpene methyl ether on proliferation and apoptosis of human gastric adenocarcinoma AGS cells
Lijing ZHANG ; Leilei ZHANG ; Wenhua HUANG ; Li GAO ; Xiaowei HUO ; Dongyu LIU ; Liyong LI ; Li CAO
Chinese Pharmacological Bulletin 2014;(8):1066-1072
Aim Toinvestigatetheeffectsoftriptonot-erpene methyl ether ( TME ) , a diterpene derived from the medicinal plant Triptergium wilfordii, on human gastric cancer AGS cell proliferation inhibition and ap-optosisinducedinvitro.Methods MTTassaywas used for screening tumor spectrum and detecting the vi-ability of AGS cells and normal human gastric epitheli-al cells GES-1 . Cell morphology was observed by light microscopy and AO / EB staining. Flow cytometry was used to detect cell apoptotic rate and cell cycle. JC-1 staining and fluorescence probe DCFH-DA were em-ployed to detect the changes of mitochondrial mem-brane potential and reactive oxygen species ( ROS ) . The effect of inhibiting AGS clonogenic survival was as-sayed by the method of plate clone formation. Western blot was used to analyse the expression of caspase-3 , caspase-8,Bcl-2andBax.Results MTTresults showed that TME exhibited significantly higher cytotox-icity to gastric cancer AGS cell line than to noncancer-ous cell line GES-1. IC50 for AGS of 48 h treatment was 23 . 85 μmol · L-1 . TME significantly inhibited colony formation and caused morphological changes in AGS cells. Annexin V-FITC / PI double staining showed the apoptotic rate increased. DCFH-DA stai-ning showed TME resulted in an increase in intracellu-lar ROS levels. Mitochondrial membrane potential de-creased after TME treatment. Western blot results showed that TME increased the proportion of Bax /Bcl-2 , with the activation of caspase-8 and caspase-3 . The broad-spectrum caspase inhibitor z-VAD-fmk pre-treatment reduced the expression of caspase-8 and caspase-3. TME enabled AGS cell cycle arrest in G0/G1phase.Conclusion TMEpossessespotenttumor selected toxicity and can induce apoptosis of AGS cells through cell cycle arrest, which is associated with Bcl-2 protein family.
4.Protective effects of CMP on 5-Fu-induced intestinal mucositis of mice
Canhong WANG ; Xiaoshan HE ; Lijing ZHANG ; Xiaowei HUO ; Dongyu LIU ; Jiayuan HAN ; Liyong LI ; Li CAO
Chinese Pharmacological Bulletin 2016;32(4):484-488,489
Aim To evaluate the mucosal-protective effects of carboxymethylpachyman( CMP) on Fluorou-racil(5-Fu)-induced mice intestinal mucositis and ex-plore its mechanisms. Methods ICR mice were as-signed randomly to four groups:normal group( n=8;re-ceiving pure water orally for 14 d) ,CMP group( n=8;200 mg·kg-1 CMP for 14 d orally),5-Fu group(n=8;25 mg·kg-1 5-Fu for 7 d,intraperitoneally( i. p. ) , and CMP+5-Fu group( n=8;200 mg·kg-1 CMP for 14 d orally and 25 mg·kg-1 5-Fu for 7 d,i. p. ). At day 14the mice were sacrificed. The intestinal propel-ling rate and the colon length were measured. ROS, GSH and IL-1βcontents,and CAT,GSH-Px activities in homogenate supernatant of PPs were measured by kits for observing the effects of CMP on mice lipid peroxida-tion and intestinal mucosal inflammatory induced by 5-Fu. Colon tissues were used for hematoxylin and eosin ( HE ) staining for the determination of the effect of CMP on mice colon histopathology, immunohistochem-istry for the protein levels of NF-κB and p-p38 . Results CMP significantly extended colon lengths,accelerate the intestinal propelling rates, reduced colonic mucosa epithelium goblet cell loss, inflammatory cells infiltra-tion,and crypt depth shallow induced by 5-Fu. CMP obviously reduced ROS and IL-1β contents, and pre-vented reductions in homogenate supernatant of PPs GSH content, CATand GSH-Px activities by 5-Fu ad-ministration,and also reduced the expression of NF-κB and p-p38 in colon tissues. However, CMP alone had no effect on the colon of normal mice. Conclusion The current study demonstrates that CMP may have sig-nificant protective effects against 5-Fu-induced intesti-nal mucositis. Its mechanism may be related to enhan-cing the antioxidant activity,anti-inflammatory and an-ti-apoptotic effects.
5.The Application and Effectiveness Evaluation of Continuing Care Based on Humanistic Care for Out Patients with Facial Plastic Surgery
Xiulin WEN ; Jieqiong LI ; Xiaowei HUO ; Jinjin LIU ; Yuan GUO ; Ting LI
Chinese Medical Ethics 2015;(5):808-811
Objective:To evaluate the effect of continuing care in facial plastic operation .Method:Select 120 patients who had aesthetic plastic surgery in our hospital during the period from January to July in 2014 and ran-domize them into the observation group (60 cases)and the control group (60 cases).The control group merely re-ceive conventional treatment .The observation group is employed continuing care based on conventional treatment . Results:The satisfaction of two groups has no significant difference a week after operation (P>0.05).Six months after operation , the satisfaction of the observation group is obviously higher than that of the control group .The difference is statistically different ( P<0 .0001 ) .The anxiety level of the observation group is obvious lower than that of the control group and the difference is Statisticallydifferent ( P<0 .0001 ) .Conclusion: Based on the hu-manistic care, carry on the continuing care could considerately improve the satisfaction of patients in clinic facial plastic surgery and is one of the most important solution of preventing medical disputes ;mean while , it is also one of the most effective stress reduction measures which makes the patients feel humanized nursing .
6.Consensus standpoints from expert panel of Chinese Society of Cardiology on AHA/ACC 2013 guideline on the treatment of blood cholesterol to reduce atherosclerotic cardiovascular risk in adults.
Xiaowei YAN ; Hong CHEN ; Wei GAO ; Jianjun LI ; Xinchun YANG ; Ping YE ; Shuyang ZHANG ; Dong ZHAO ; Jianhua ZHU ; Yong HUO
Chinese Journal of Cardiology 2014;42(4):275-276
7.Rhodococcus equi distribution in the soil environment of horses in Inner Mongolia,China——An investigation report
Zhuang DING ; Takai SHINJI ; Madarame HIROO ; Shuang CHANG ; Hainan HUANG ; Xiaowei HUO ; Minghua GAO ; Zhongtian TAN ; Shuangcheng GAO ; Hatori FUMIKO ; Sasaki YUKAKO ; Kakuda TSUTOMU ; Tsubaki SHIRO
Chinese Journal of Veterinary Science 2008;28(1):40-44
Little is known about the distribution of Rhodococcus equi in the soil environment of native horses in China. One hundred and eight soil samples were collected from native-horse farms in the Hulun Beier grasslands of eastern Mongolia, the Xilin Goler grasslands of southern Mongolia, and Tongliao city in Inner Mongolia of China for investigating the distribution of R. equi in these regions. The isolation rates of R. equi from soil samples from the Hulun Beier and Xilin Goler grasslands ranged from 25.9% to 30.0%. In contrast, isolation rates from soil samples from Tongliao city was as high as 82.3% and the mean number of R. equi in soil samples from Tongliao city was 10 times more than those of samples from the grasslands. The 488 isolates were examined using PCR for the presence of genes that encode virulence-associated 15 000-17 000 antigen protein (VapA) and the 20 000 antigen protein (VapB). All isolates were negative for virulence-associated proteins. Plasmid profiles of these avirulent isolates showed that cryptic plasmids of various sizes were present with an incidence of 13.3% to 21.5%. The results of the present study contrast with those of our recent study, in which we reported that R. equi was absent from Mongolian horses in Ulaanbaatar, Mongolia. It is suggested that the difference between the results of these two studies is due to the mobile pasturing system in Mongolia and nonmobile pasturing system in Inner Mongolia.
8. Primary carcinoid of ovary: a clinicopathologic analysis of 17 cases
Ran ZHU ; Xiaowei XUE ; Yufeng LUO ; Huanwen WU ; Zhen HUO
Chinese Journal of Pathology 2018;47(5):339-343
Objective:
To study the clinicopathologic feature, diagnostic strategy and prognostic significance of primary carcinoid of the ovary (PCOTO).
Methods:
A series of 17 patients previously diagnosed as PCOTO at Department of Pathology, Peking Union Medical College Hospital during the period from 2002 to 2017 were evaluated with clinical data analysis, histopathology and immunohistochemistry, and the patients were followed up and the relevant literatures were reviewed.
Results:
The age of patients ranged from 24 to 64 years (mean, 42 years). Fourteen patients were found a pelvic mass for a health check-up, and only 3 patients presented with pain in the lower abdomen. The left ovary was involved initially in ten patients, and the right in seven. The major axis of the tumor ranged from 2 to 14 cm. The surface of these tumors was smooth. Seven of the tumors were solid-cystic, 6 were cystic and 4 were solid. Under light microscope, 6 cases were diagnosed as strumal carcinoid, 4 cases were insular carcinoid, 4 cases were trabecular carcinoid, 3 cases were insular and trabecular mixed type carcinoid and 1 case was mucinous carcinoid. The mitotic figures were no more than 1/10 HPF.There were 11 cases complicated with other ovarian tumors, including 10 cases with teratoma, and 1 case with mucinous cystic adenoma. The paraffin-embedded tissues of all cases showed immunoreactivity for NSE and Syn, and the positive propotion of CgA was 10/17. TTF1 was positive in thyroid follicles and negative in strumal carcinoid. The positive index of Ki-67 was no more than 2%. Follow-up of 13 to 188 months showed 16 patients without recurrence and 1 patients were loss to follow-up.
Conclusions
PCOTO is very rare. Most of the patients are found with a mass during health check-up in unilateral ovary and without obvious clinical symptoms. Histologically, the tumor always exists with other ovarian tumors, including teratoma and mucinous cystic adenoma, with relatively good prognosis.
9. The application of artificial dermis in repairing partial fingernail bed defect
Shaohua LIU ; Zhanbei MA ; Hetong YU ; Liang MA ; Yuechuang LIANG ; Yu WU ; Dongyue LI ; Chong NAN ; Huanhuan ZHAO ; Xiaowei REN ; Lie HUO
Chinese Journal of Plastic Surgery 2018;34(11):955-958
Objective:
To explore the clinical effect of artificial dermis on partial nail bed defects.
Methods:
From May 2013 to September 2016, 16 cases of 21 fingers with partial nail bed defect and bone exposure received primary repair with artificial dermis after debridement. Then the outer dressing was changed 3 days after the operation, and the dressing was changed once every 3 to 5 days. The pink dermis like tissue was placed on the wound surface 2 to 3 weeks after the operation. The wound surface was naturally epithelialized.
Results:
All the fingers were survived after artificial dermis repair, and the fingertip surface of nail bed defect was epithelialized naturally and healed well. Postoperative follow-up was 5 to 16 months, with an average of 8.2 months. The growth of fingernails was smooth with symmetrical appearance, natural toughness, good adhesion between the nail body and the nail bed, and no pain or hyperesthesia at the tip. Results of curative effect evaluation showed excellent in 18 fingers, good in 3 fingers. Results of patient′s satisfaction with postoperative finger appearance showed very satisfactory in 13 cases and satisfactory in 3 cases.
Conclusions
The method of repairing partial defect of nail bed using artificial dermis is simple with minimal damage and a good clinical effect.
10.Analysis of codon usage patterns in Bupleurum falcatum chloroplast genome.
Mengqi GAO ; Xiaowei HUO ; Liting LU ; Mengmeng LIU ; Gang ZHANG
Chinese Herbal Medicines 2023;15(2):284-290
OBJECTIVE:
In order to distinguish the traditional Chinese medicine Bupleurum falcatum and its adulterants effectively and develop a better understanding of the factors affecting synonymous codon usage, codon usage patterns of chloroplast genome, we determine the complete chloroplast (cp) genome of B. falcatum and clarify the main factors that influence codon usage patterns of 78 genes in B. falcatum chloroplast genome.
METHODS:
The total genomic DNA of fresh leaves from a single individual of B. falcatum was extracted with EASYspin plus Total DNA Isolation Kit and 2 μg genome DNA was sequenced using Illumina Hiseq 2500 Sequencing Platform. The cp genome of B. falcatum was reconstructed with MITObim v1.8 and annotated in the program CPGAVAS2 with default parameters. Python script and Codon W were used to calculate the codon usage bias parameters.
RESULTS:
The full length of B. falcatum cp genome was 155 851 bp, 132 different genes were annotated in this cp genome containing 80 protein-coding genes, 30 tRNA genes, and four rRNA genes. The codon usage models tended to use A/T-ending codons. The neutrality plot, ENC plot, PR2-Bias plot and correspondence analysis showed that both compositional constraint under selection and mutation could affect the codon usage models in B. falcatum cp genome. Furthermore, three optimal codons were identified and most of these three optimal codons ended with G/U.
CONCLUSION
The cp genome of B. falcatum has been characterized and the codon usage bias in B. falcatum cp genome is influenced by natural selection, mutation pressure and nucleotide composition. The results will provide much more barcode information for species discrimination and lay a foundation for future research on codon optimization of exogenous genes, genetic engineering and molecular evolution in B. falcatum.