1.Screening,validation,and functional analysis of Brucella secretory BspE interac-ting host proteins
Shuanghong YIN ; Xiaoyu DENG ; Hongyan LIU ; Haixiao WANG ; Caixia YI ; Yincui LI ; Xin SUN ; Shuli WANG ; Jihai YI ; Junbo ZHANG
Chinese Journal of Veterinary Science 2024;44(7):1438-1447,1457
In order to explore the role of BspE protein in Brucella infection,yeast two-hybrid tech-nique was used to screen host cell proteins that interact with BspE protein.The constructed BspE recombinant plasmid pGBKT7-BspE was used as bait plasmid to hybridize with the RAW264.7-cD-NA library of mouse mononuclear macrophages by yeast two-hybridization technique.The positive clones were extracted by plasmid,sequenced and co-immunoprecipitation to determine the host cell proteins that could interact with BspE.The subcellular localization of BspE proteins was analyzed by confocal laser microscopy.The physical and chemical properties,protein structure and function of BspE interacting proteins were analyzed by bioinformatics.The siRNA for one of the BspE inter-acting proteins was synthesized,the expression of its gene was silenced in HEK293T cells,and the silenced cells was infected with Brucella M5-90 and the number of intracellular bacteria was coun-ted.The results showed that the decoy plasmid pGBKT7-BspE was successfully constructed,and the plasmid could express BspE protein in yeast.Eight positive clones were obtained from the host cell genome library by yeast two-hybridization.The positive clones were identified as RBM27 and PCBP1 by sequencing,backcross and co-immunoprecipitation.Bioinformatics was used to predict the cell location,protein structure and amino acid composition of RBM27 and PCBP1.After siRNA interference,the expression level of PCBP1 was significantly decreased and the amount of M5-90 in the cell was increased.Brucellosis secreted protein BspE interacts with host proteins RBM27 and PCBPl,and PCBP1 negatively regulates the proliferation of Brucellosis.
2.Aging-Induced Endothelial Glycocalyx Alteration and Vascular Dysfunction
Zihang ZHANG ; Duolan GAO ; Xinyuan DAI ; Tian LIU ; Minghan LI ; Xiaoyan DENG ; Hongyan KANG
Journal of Medical Biomechanics 2024;39(2):368-374
The endothelial glycocalyx(EG)is a polyglycoprotein complex present on the internal vascular surface,and its impairment is associated with the progression of multiple diseases,including atherosclerosis,stroke,sepsis,diabetes,kidney disease,hypertension,and lung edema.Therefore,glycocalyx health can be used as a biomarker to evaluate vascular health.Aging leads to dysfunctional changes in the glycocalyx;for example,its thickness decreases,and the genes of enzymes involved in its synthesis and digestion are dysregulated.As a natural barrier to the vascular system,age-related glycocalyx disruption is associated with vascular dysfunction,including impairment of vascular contraction and dilation,enhancement of permeability,dysregulation of inflammatory and immune reactions,and imbalance of anticoagulation and thrombin.From the perspective of'structure determines function'studies on the changing regularity of the thickness,components,microstructure,and mechanical properties of EG with aging and its relationship with vascular dysfunction are of great significance for the prevention,diagnosis,and treatment of atherosclerosis and other age-related cardiovascular diseases.
3.Application of Multimodal Ultrasound in Diagnosis,Typing and Staging of Primary Nodal Diffuse Large B-Cell Lymphoma
Hongyan DENG ; Wenjuan LU ; Yasu ZHOU ; Ya YUAN ; Xinhua YE
Chinese Journal of Medical Imaging 2024;32(11):1111-1117
Purpose To investigate the application value in diagnosis,typing and staging of multimodal ultrasound in primary nodal diffuse large B-cell lymphoma(N-DLBCL).Materials and Methods A total of 96 patients(96 nodes)with a pathological diagnosis of N-DLBCL(51 nodes)or benign lymph nodes(45 nodes)in the First Affiliated Hospital of Nanjing Medical University from October 2020 to June 2022 were enrolled,retrospectively.All these patients were examined by high-frequency ultrasound,shear wave elastography(SWE)and contrast-enhanced ultrasonography(CEUS).The differences among the three ultrasound imaging methods in differentiating N-DLBCL from benign lymph nodes,typing and staging of N-DLBCL were analyzed.Receiver operating characteristic curve were drawn to obtain the diagnostic thresholds of SWE and CEUS quantitative indexes,and the efficacy of the different examination methods were assessed.Results Univariate analysis demonstrated 14 parameters were significantly different between the N-DLBCL and benign lymph nodes(χ2=12.289-32.934,all P<0.05).Multivariate analysis showed the difference in long diameter/short diameter(OR=11.205,P=0.012),cortical echo(OR=10.367,P=0.002),maximum elasticity(OR=0.180,P=0.014),peak time(OR=0.111,P=0.025),peak intensity-basic intensity(OR=0.061,P=0.002)and enhancement distribution(OR=0.065,P=0.001)were statistically significant between the two groups.There were statistically significant differences of maximum elasticity(χ2=5.299,P=0.021)between germinal center and non-germinal center N-DLBCL groups.There were statistically significant differences in blood flow pattern(χ2=9.663,P=0.017),arrival time(χ2=2.851,P=0.034)and peak time(χ2=6.702,P=0.018)between the limited and advanced N-DLBCL.The area under the curve for the diagnosis of N-DLBCL and benign lymph nodes by high-frequency ultrasound,SWE,CEUS and multimodal ultrasound were 0.754,0.839,0.875 and 0.963,respectively.Conclusion Multimodal ultrasound has high application value in differential diagnosis,classification and staging of N-DLBCL,and can provide basis for diagnosis and treatment of N-DLBCL.
4.Effects of 2 650 MHz radiofrequency radiation on the behavior and hippocampal neurotransmitter release of mice
Yujie LIU ; Jun WANG ; Keqin LI ; Chenxu CHANG ; Ying LIU ; Hongyan ZUO ; Yang LI ; Hong YANG ; Yanhui HAO ; Hua DENG
Chinese Journal of Radiological Medicine and Protection 2024;44(5):354-360
Objective:To investigate the effects of 2 650 MHz radiofrequency (RF) exposure on the behavior and neurotransmitter release of mice.Methods:Adult male C57BL/6N mice were divided into a normal control (CON) group and a radiofrequency radiation (RFR) group using the random number table method. The mice in the RFR group were subjected to single-dose whole-body exposure to a uniform 2 650 MHz RF electromagnetic field for 3 h. During the RF exposure, the field strength in the effective working area of the RF radiation platform was measured using an electromagnetic radiation analyzer, and the changes in the anal temperature of the mice were monitored using an optical fiber thermometer. Moreover, the changes in the cognition, social interaction, and emotion of the mice were determined through the new object recognition test, social preference test, and open field test. Finally, the changes in the hippocampal neurotransmitter release levels of the mice were detected using microdialysis sampling and mass spectrometry, and the changes in the hippocampal tissue structure and ultrastructure were observed via microscopy.Results:Under the test conditions, RF radiation improved the anal temperature of the mice, with a maximum increasing amplitude of 0.61℃, falling within the range of thermal safety. The mice in the RFR group experienced a significant decrease in the frequency and time for exploring new objects ( t=4.50, 2.53, P < 0.05) in the new object recognition test, a significant decrease in the frequency ( t=0.08, P<0.01) and time ( t=0.03, P<0.05) for exploring other mice in the social preference test, and no significant change in the frequency and time for exploring the central area ( P > 0.05) in the open field test. Compared to the CON group, the RFR group showed an increase in the release of 5-hydroxytryptamine (5-HT) ( t=-2.56, P < 0.05) and a decrease in the release of acetylcholine (ACh) ( t=2.21, P < 0.05), no significant difference in the release of glutamate (Glu) and γ-aminobutyric acid (GABA) ( P > 0.05), and no evident damage to the hippocampal tissue and structure and synaptic ultrastructure. Conclusions:2 650 MHz RF radiation may induce cognitive impairment and abnormal social preference in mice, which is attributed to neuronal dysfunctions and neurotransmitter release disorders under RF exposure.
5.Application and evaluation of management mode for the use of antimicrobial drugs in county-level medical communities led by anti-infective clinical pharmacists
Xiaoqin DENG ; Chi ZHAO ; Zhaohong LI ; Hongyan YAN ; Dongfang SHEN ; Helang TAN ; Mingzhong JIANG ; Nanjun DENG
China Pharmacy 2024;35(1):95-100
OBJECTIVE To provide reference for improving the rational use of antimicrobial drugs in primary township medical institutions. METHODS Based on the county prescription pre-review center, a team led by anti-infective clinical pharmacists constructed the management mode for the use of antimicrobial drugs in county-level medical communities with clinical pharmacists as the main team by finding out the main problems in the use of antimicrobial drugs in primary township medical institutions, providing feedback on the problems, organizing relevant training for the problems, improving the customization rules of the prescription pre-review software, implementing the automatic interception and pharmacist online prescription review and other measures. Data on the use of antimicrobial drugs were collected and compared in the 15 primary township medical institutions between January-June in 2022 (before the implementation of the mode) and January-June in 2023 (after the implementation of the mode). RESULTS Compared with before the implementation of the mode, the utilization rate of antimicrobial drugs in outpatients of primary township medical institutions decreased from 24.97% before the implementation of the mode to 19.39% after the implementation of the mode; the utilization rate of antimicrobial injection in outpatients decreased from 66.10% to 46.80%; the utilization rate of intravenous drip of antimicrobial drugs in outpatients decreased from 52.33% to 40.35%; the rates of combined use of antimicrobial drugs in outpatients decreased from 12.70% to 8.19%; the reasonable rate of antimicrobial prescribing in outpatients increased from 55.28% to 73.93%. After the implementation of the mode, the proportion of antimicrobial prescriptions for each diagnosis was basically the same as before; the defined daily dose system (DDDs) and proportion of a few antimicrobial drugs changed compared with before according to the anatomical therapeutic chemical classification of drugs, among which DDDs of lincomycin, gentamicin and other drugs declined significantly; DDDs of antimicrobial drugs for each classification was basically the same as before according to AWaRe classification. CONCLUSIONS The management mode of the use of antimicrobial drugs led by anti-infective clinical pharmacists is constructed in the prescription pre-reviewing center of county-level medical communities, which can effectively improve the rational use of antimicrobial drugs in the primary township medical institutions.
6.Mechanisms by which Mettl3 regulates pericyte-myofibroblast transdifferentiation through PI3K/AKT signaling pathway
Yi DENG ; Yan WANG ; Pingping HE ; Jiao LI ; Weiwei LIU ; Jinsong YUAN ; Hongyan ZHAO ; Zhijiang LIU ; Changyin SHEN ; Bei SHI
Chinese Journal of Cardiology 2024;52(7):814-826
Objective:To investigate the role and underlying mechanisms of methyltransferase (Mettl) 3 in the process of angiotensin Ⅱ (Ang Ⅱ)-induced pericyte-to-myofibroblast transdifferentiation and renal fibrosis.Methods:C57BL/6J mice were used, in cell experiments, mouse renal pericytes were isolated and cultured using magnetic bead sorting. These pericytes were then induced to transdifferentiate into myofibroblasts with 1×10 6 mmol/L Ang Ⅱ, which was the Ang Ⅱ group, while pericytes cultured in normal conditions served as the control group. Successful transdifferentiation was verified by immunofluorescence staining, Western blotting, and real-time reverse transcription PCR (RT-qPCR) for α-smooth muscle actin (α-SMA). The levels of m6A modifications and related enzymes (Mettl3, Mettl14), Wilms tumor 1-associated protein (WTAP), fat mass and obesity protein (FTO), ALKBH5, YTHDF1, YTHDF2, YTHDC1, YTHDC2, YTHDC3 were assessed by Dot blot, RT-qPCR and Western blot. Mettl3 expression was inhibited in cells using lentivirus-mediated Mettl3-shRNA transfection, creating sh-Mettl3 and Ang Ⅱ+sh-Mettl3 groups, while lentivirus empty vector transfection served as the negative control (Ang Ⅱ+sh-NC group). The impact of Ang Ⅱ on pericyte transdifferentiation was observed, and the expression of downstream phosphatidylinositol 3-kinase (PI3K)/AKT signaling pathway proteins, including PI3K, AKT, phosphorylated AKT at serine 473 (p-AKT (S473)), and phosphorylated AKT at threonine 308 (p-AKT (T308)), were examined. PI3K gene transcription was inhibited by co-culturing cells with actinomycin D, and the half-life of PI3K mRNA was calculated by measuring residual PI3K mRNA expression over different co-culture time. The reversibility of Mettl3 inhibition on Ang Ⅱ-induced pericyte-to-myofibroblast transdifferentiation was assessed by adding the AKT activator SC79 to the Ang Ⅱ+sh-Mettl3 group. In animal experiments, mice were divided into these groups: sham group (administered 0.9% sterile saline), Ang Ⅱ group (infused with Ang Ⅱ solution), sh-Mettl3 group (injected with Mettl3 shRNA lentivirus solution), Ang Ⅱ+sh-Mettl3 group (infused with Ang Ⅱ solution and injected with Mettl3 shRNA lentivirus solution), and Ang Ⅱ+sh-Mettl3+SC79 group (administered Ang Ⅱ solution and Mettl3 shRNA lentivirus, with an additional injection of SC79). Each group consisted of six subject mice. Blood pressure was measured using the tail-cuff method before and after surgery, and serum creatinine, urea, and urinary albumin levels were determined 4 weeks post-surgery. Kidney tissues were collected at 28 days and stained using hematoxylin-eosin (HE) and Masson′s trichrome to assess the extent of renal fibrosis. Results:Primary renal pericytes were successfully obtained by magnetic bead sorting, and intervened with 1×10 6 mmol/L Ang Ⅱ for 48 hours to induce pericyte-to-myofibroblast transdifferentiation. Dot blot results indicated higher m6A modification levels in the Ang Ⅱ group compared to the control group ( P<0.05). RT-qPCR and Western blot results showed upregulation of Mettl3 mRNA and protein levels in the Ang Ⅱ group compared to the control group (both P<0.05). In the Ang Ⅱ+sh-Mettl3 group, Mettl3 protein expression was lower than that in the Ang Ⅱ group, with reduced expression levels of α-SMA, vimentin, desmin, fibroblast agonist protein (FAPa) and type Ⅰ collagen (all P<0.05). Compared to the control group, PI3K mRNA expression level was elevated in the Ang Ⅱ group, along with increased p-AKT (S473) and p-AKT (T308) expressions. In the Ang Ⅱ+sh-Mettl3 group, PI3K mRNA expression and p-AKT (S473) and p-AKT (T308) levels were decreased (all P<0.05). The half-life of PI3K mRNA was shorter in the Ang Ⅱ+sh-Mettl3 group than that in the Ang Ⅱ+sh-NC group (2.34 h vs. 3.42 h). The ameliorative effect of Mettl3 inhibition on Ang Ⅱ-induced pericyte-to-myofibroblast transdifferentiation was reversible by SC79. Animal experiments showed higher blood pressure, serum creatinine, urea, and 24-hour urinary protein levels, and a larger fibrosis area in the Ang Ⅱ group compared to the sham group (all P<0.05). The fibrosis area was smaller in the Ang Ⅱ+sh-Mettl3 group than that in the Ang Ⅱ group ( P<0.05), but increased again upon addition of SC79. Conclusion:Mettl3-mediated RNA m6A epigenetic regulation is involved in Ang Ⅱ-induced pericyte-to-myofibroblast transdifferentiation and renal fibrosis, potentially by affecting PI3K stability and regulating the PI3K/AKT signaling pathway.
7.miR-34b-3p Inhibition of eIF4E Causes Post-stroke Depression in Adult Mice.
Xiao KE ; Manfei DENG ; Zhuoze WU ; Hongyan YU ; Dian YU ; Hao LI ; Youming LU ; Kai SHU ; Lei PEI
Neuroscience Bulletin 2023;39(2):194-212
Post-stroke depression (PSD) is a serious and common complication of stroke, which seriously affects the rehabilitation of stroke patients. To date, the pathogenesis of PSD is unclear and effective treatments remain unavailable. Here, we established a mouse model of PSD through photothrombosis-induced focal ischemia. By using a combination of brain imaging, transcriptome sequencing, and bioinformatics analysis, we found that the hippocampus of PSD mice had a significantly lower metabolic level than other brain regions. RNA sequencing revealed a significant reduction of miR34b-3p, which was expressed in hippocampal neurons and inhibited the translation of eukaryotic translation initiation factor 4E (eIF4E). Furthermore, silencing eIF4E inactivated microglia, inhibited neuroinflammation, and abolished the depression-like behaviors in PSD mice. Together, our data demonstrated that insufficient miR34b-3p after stroke cannot inhibit eIF4E translation, which causes PSD by the activation of microglia in the hippocampus. Therefore, miR34b-3p and eIF4E may serve as potential therapeutic targets for the treatment of PSD.
Animals
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Mice
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Depression
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Eukaryotic Initiation Factor-4E/metabolism*
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MicroRNAs/metabolism*
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Neurons/metabolism*
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Stroke/metabolism*
8.Multicenter retrospection and analysis of influencing factors on blood transfusion in patients with extensive burns
Deqing DUAN ; Yong CHEN ; Hong'ao DENG ; Shiqiang HU ; Yuangui MAO ; Dewu LIU ; Chunmao HAN ; Qinglian XU ; Hongyan ZHANG
Chinese Journal of Burns 2023;39(11):1047-1056
Objective:To retrospect the blood transfusion status of patients with extensive burns in multiple centers and analyze its influencing factors.Methods:A retrospective case series study was conducted. Clinical data of 455 patients with extensive burns who met the inclusion criteria and were admitted to the burn centers of 3 hospitals from January 2016 to June 2022 were collected, including 202 patients from the First Affiliated Hospital of Nanchang University, 179 patients from the Second Affiliated Hospital of Zhejiang University School of Medicine, and 74 patients from the First Affiliated Hospital of Anhui Medical University. The following data were collected from patients during their hospitalization, including infusion of red blood cells, plasma, and platelets during hospitalization; age, gender, body mass index, combined underlying diseases, cause of injury, time of admission after injury, type of admission, total burn area, full-thickness burn area, combination of inhalation injury, combination of other trauma, and combination of pulmonary edema; the blood lactic acid, serum creatinine, total bilirubin, and albumin values within 24 h of admission; combination of bloodstream, wound, lung, and urinary tract infection, and combination of sepsis; the number of escharectomy or tangential excision and skin grafting surgery (hereinafter referred to as surgery) and total surgical blood loss volume; occurrence of hemoglobin<70 g/L, admission to intensive care unit (ICU), conduction of mechanical ventilation and continuous renal replacement therapy (CRRT), length of hospital stay, and prognosis were recorded. In 602 surgeries of patients within 14 days after injury, data including area of escharectomy or tangential excision and skin graft harvesting, duration of operation, and surgical blood loss volume per surgery, operation site, and use of tourniquet and wound graft were collected. Data were statistically analyzed with Mann-Whitney U test, Kruskal-Wallis H test, and Spearman correlation analysis. Combined with the results of single factor analysis and clinical significance, multiple linear regression analysis was performed to screen the independent influencing factors of red blood cell infusion volume and plasma infusion volume, as well as blood loss volume per surgery. Results:During the whole hospitalization period, 437 (96.0%) patients received blood transfusion therapy, including 435 (95.6%) patients, 410 (90.1%) patients, and 73 (16.0%) patients who received transfusion of plasma, red blood cells, and platelets, respectively. The patients were mainly male, aged 18 to 92 years. There were statistically significant differences in the plasma infusion volume among patients with different combination of underlying disease, combination of inhalation injury, combination of other trauma, combination of pulmonary edema, combination of bloodstream infection, combination of wound infection, combination of lung infection, combination of urinary tract infection, combination of sepsis, occurrence of hemoglobin value <70 g/L, admission to ICU, conduction of mechanical ventilation, and conduction of CRRT (with Z values of -2.06, -4.67, -2.11, -6.13, -9.56, -4.93, -8.08, -4.78, -9.12, -6.55, -9.37, -11.46, and -7.17, respectively, P<0.05). The total burn area, full-thickness burn area, blood lactic acid value within 24 h of admission, serum creatinine value within 24 h of admission, albumin value within 24 h of admission, number of surgeries, and total surgical blood loss volume were correlated with the plasma infusion volume of patients (with r values of 0.39, 0.51, 0.14, 0.28, -0.13, 0.47, and 0.56, respectively, P<0.05).There were statistically significant differences in the red blood cell infusion volume among patients with different gender, combination of inhalation injury, combination of other trauma, combination of pulmonary edema, combination of bloodstream infection, combination of wound infection, combination of lung infection, combination of urinary tract infection, combination of sepsis, occurrence of hemoglobin value <70 g/L, admission to ICU, conduction of mechanical ventilation, and conduction of CRRT (with Z values of -2.00, -4.34, -3.10, -4.22, -8.24, -7.66, -8.62, -4.75, -7.42, -9.36, -6.12, and -8.31, -6.64, respectively, P<0.05). The age, total burn area, full-thickness burn area, blood lactic acid value within 24 h of admission, serum creatinine value within 24 h of admission, total bilirubin value within 24 h of admission, number of surgeries, and total surgical blood loss volume were correlated with the red blood cell infusion volume of patients (with r values of 0.12, 0.22, 0.49, 0.09, 0.18, 0.13, -0.15, 0.69, and 0.77, respectively, P<0.05). Combined underlying diseases, full-thickness burn area, combined pulmonary edema, serum creatinine value within 24 h of admission, combined sepsis, conduction of CRRT, number of surgeries, and total surgical blood loss volume were the independent influencing factors for plasma infusion volume during hospitalization in patients with extensive burns (with standardized regression coefficients of 0.09, 0.16, 0.12, 0.07, 0.11, 0.15, 0.31, and 0.26, respectively, P<0.05). Female, full-thickness burn area, serum creatinine value within 24 h of admission, combined sepsis, occurrence of hemoglobin value <70 g/L, conduction of CRRT, and total surgical blood loss volume were the independent influencing factors for red blood cell infusion volume during hospitalization in patients with extensive burns (with standardized regression coefficients of 0.10, 0.12, 0.10, 0.11, 0.05, 0.19, and 0.54, respectively, P<0.05). There were statistically significant differences in blood loss volume per surgery of patients with different surgical site and wound graft (with Z values of -2.54 and -2.27, respectively, P<0.05). The area of escharectomy or tangential excision and skin graft harvesting and duration of operation were correlated with the blood loss volume per surgery of patients (with r values of 0.40 and 0.21, respectively, P<0.05). The area of escharectomy or tangential excision and skin graft harvesting, duration of operation, and active wound grafts were the independent influencing factors for blood loss volume per surgery of patients with extensive burns (with standardized regression coefficients of 0.41, 0.16, and 0.12, respectively, P<0.05). Conclusions:The major factors influencing blood transfusion status in patients with extensive burns are female, combined underlying diseases, full-thickness burn area, serum creatinine value within 24 h of admission, combined pulmonary edema, occurrence of hemoglobin value <70 g/L, combined sepsis, conduction of CRRT, number of surgery, and total surgical blood loss volume. In addition, the area of escharectomy or tangential excision and skin graft harvesting, duration of operation, and active wound grafts indirectly affect the patient's blood transfusion status by affecting the blood loss volume per surgery.
9.Analysis of HPV infection and subtypes in 60 354 patients in Changsha
Ping LI ; Shaoqiang CHEN ; Changjuan DENG ; Jing LI ; Hui ZHOU ; Hongyan WANG ; Mengzhu LI ; Xiaoying KUANG ; Xiaobing XIE
Chinese Journal of Laboratory Medicine 2023;46(4):360-366
Objective:Analysis of subtype distribution characteristics of human papillomavirus (HPV) infection, pathological findings of HPV-positive patients operated colposcopy and cervical histopathological examination in Changsha from 2020 to 2022.Methods:Retrospective analysis of HPV infection status of 60 354 patients was treated in the First Affiliated Hospital of Hunan University of Chinese Medicine from Jan. 2020 to Oct. 2022. They were divided into<25 years old group (3 250 cases), 25-34 years old group (19 406 cases), 35-44 years old group (17 297 cases), 45-54 years old group (13 104 cases), 55-64 years old group (4 793 cases) and≥65 years old group (2 504 cases). Cervical exfoliated cells were collected from women and specimen of lesion site were collected from men. HPV genotyping detection used polymerase chain reaction (PCR) and flow fluorescence hybridization, which could detect 27 HPV genotypes. Analyze the distribution of HPV subtypes in patients of different age groups and different treatment departments. Analyze the relationship between the infection of HPV subtypes and the outcome of 224 patients with HPV positive who also underwent colposcopy and cervical histopathology. Pearson correlation test was used for correlation analysis, and two-tailed P<0.05 was statistically significant; Pearson chi-square test or continuous correction chi-square test was used to compare the rates between groups, and two-tailed P<0.05 was statistically significant; Chi-square split test was used to compare multiple sample rates. The standard test was corrected according to the number of groups and then compared. The difference was statistically significant when the two-tailed P value was less than the corrected standard test. Results:The overall positive rate of HPV was 16.4% (9 909/60 354). Among all HPV positive patients, affection of single type HPV accounted for 75.5% (7 479/9 909) and affection of multi-type HPV accounted for 24.5% (2 430/9 909). The top six HPV types with infection rates from high to low were: HPV52 22.8% (2 256/9 909), HPV58 11.1% (1 097/9 909), HPV53 10.5% (1 045/9 909), HPV16 9.0% (890/9 909), HPV61 7.8% (774/9 909) and HPV6 7.6% (750/9 909). The difference of overall infection rate between different age groups was statistically significant (χ 2=536.90, P<0.001). The HPV infection rate was the highest in the<25 years old group, 30.1% (978/3 250), and it is higher than the age groups of 25-34 years old, 15.6% (3 035/19 406), group of 35-44 years old, 14.2% (2 464/17 297), group of 45-54 years old, 16.1% (2 115/13 104), group of 55-64 years old, 19.1% (915/4 793) and group of≥65 years old, 16.1% (402/2 504), with statistical significance( P<0.001 respectively). The most common types of HPV infection were type 6, 16.7% (378/2 266) and type 11, 10.5% (239/2 266) in dermatology clinics. The positive rate of HPV16 accounted for 12/17 in cervical intraepithelial neoplasia (CIN) 3 and cervical cancer cases, it was significantly higher than that in the inflammatory cases, 25.0% (35/140) and CIN1 cases, 23.0% (11/48), the statistical values were χ 2=15.02, P<0.001 and χ 2=12.48, P<0.001, respectively, all of which were statistically significant. Conclusions:HPV infection rate is highest in young people under 25 years old. Low-risk HPV6 and 11 are the main types of skin and venereal diseases. Among the total cases, high-risk HPV52, 58 and 53 are the most common types of infection with women in Changsha. However, high-grade cervical intraepithelial neoplasia and cervical cancer are more closely related to HPV16 infection.
10. Research on the sensitivity of Streptococcus agalactiae to omadacycline
ZOU Fanlu ; SHI Yiyi ; YU Zhijian ; PAN Weiguang ; WANG Hongyan ; CHENG Hang ; DENG Xiangbin ; XIONG Yanpeng
China Tropical Medicine 2023;23(9):965-
Abstract: Objective To investigate the antimicrobial activity of omadacycline (OMC) against clinical Streptococcus agalactiae (GBS) isolates, as well as its relationship with biofilm formation, resistance genes and virulence genes. Methods A total of 136 strains of Streptococcus agalactiae isolated from Shenzhen Nanshan People's Hospital between 2015 to 2020. The minimum inhibitory concentration (MIC) of OMC against Streptococcus agalactiae was determined by broth microdilution. Crystal violet staining was used to detect the biofilm formation ability of GBS. Resistance genes (tetM, tetO, tetK, ermB, OptrA) and virulence genes (cpsⅢ, bca, fbsA, cpsA, scpB) were investigated by polymerase chain reaction (PCR). Results Among the 136 clinical isolates of GBS, 20 strains (14.7%) were resistant to OMC, 64 (47.1%) were intermediate, and 52 (38.2%) were sensitive. Fifty-seven strains (41.9%) were biofilm-positive, 20 of which (35.1%) were sensitive to OMC. Seventy-nine strains (58.1%) were biofilm-negative, 32 of which (40.5%) were susceptible to OMC. There was a statistically significant difference in the sensitivity rates between the two groups of strains (χ2=63.062, P<0.001), but there was no significant difference in the sensitivity of OMC among the biofilm-positive strains (Fisher's exact test, P=0.824). The resistance rates of tetM, tetO, ermB and OptrA positive strains were higher than those of negative strains, while tetK was opposite. The presence of tetM (Z=0.815, P=0.415), tetO (Z=0.151, P=0.88), tetK (Z=0.567, P=0.571), ermB (Z=1.198, P=0.231) resistance genes in Streptococcus agalactiae had no significant impact on the sensitivity of OMC. However, the presence of the OptrA resistance gene showed a statistically significant effect on the sensitivity of OMC (Z=2.913, P=0.004). The virulence factors cpsⅢ, bca, fbsA, cpsA and scpB were all detected at a rate higher than 50%. The presence of the virulence genes cpsⅢ (Z=0.222, P=0.824), bca (Z=0.141, P=0.888), fbsA (Z=0.813, P=0.416), and cpsA (Z=1.615, P=0.106) in Streptococcus agalactiae had no significant impact on the sensitivity of OMC. However, there was a significant inter-group difference in the scpB virulence gene (Z=2.844, P=0.004), but the rank mean values and resistance rates of scpB-positive strains were lower than those of the negative strains. Conclusions The formation of biofilm in Streptococcus agalactiae reduces its sensitivity to OMC, but there was no significant difference in the sensitivity to OMC among the biofilm-positive strains. The presence of resistance genes tetM, tetO, tetK, ermB, and virulence genes cpsⅢ, bca, fbsA, cpsA, scpB in Streptococcus agalactiae is not associated with OMC resistance, but the presence of the resistance gene OptrA is correlated with OMC resistance..

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