1.Zuoguiwan Prevent Decreased Ovarian Reserve in Prenatal Stressed Offspring Rats via TLR4/NF-κB p65 Signaling Pathway
Yinjuan LYU ; Weiheng ZHANG ; Siyi CHEN ; Wenjing TIAN ; Jian GONG ; Yuhao MENG ; Huan HE ; Xiaocui JIANG ; Min ZHAO ; Min XIAO
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(17):67-76
ObjectiveTo observe the effect of Zuoguiwan on ovarian reserve in the female offspring rat model of prenatal stress (PS) and explore the mechanism based on Toll-like receptor 4/nuclear factor-κB p65 (TLR4/NF-κB p65) signaling pathway. MethodThirty-two pregnant rats were prepared and randomized into four groups (n=8): control, model, Zuoguiwan (18.9 mg·kg-1), and vitamin E (1.44 mg·kg-1). Except the control group, the other three groups were subjected to chronic unpredictable mild stress (CUMS) from day 11 of pregnancy, and the modeling was accompanied by gavage with corresponding drugs until delivery. The PS model was evaluated by the sucrose preference test, open field test, and serum corticosterone (CORT) level. The estrous cycle was monitored and the morphological changes in the ovarian tissue were observed. The serum levels of estradiol (E2), luteinizing hormone (LH), follicle-stimulating hormone (FSH), and anti-Mullerian hormone (AMH) in the 75-day-old offspring rats were measured by enzyme-linked immunosorbent assay (ELISA) to evaluate the ovarian reserve. The ovary and uterus indices were calculated. The serum levels of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) were measured by enzyme-linked immunosorbent assay (ELISA). The morphology of the ovarian tissue in the offspring on the day of birth and day 75 after birth was observed by hematoxylin-eosin staining. The transport of NF-κB p65 to the nucleus in the ovaries of the 75-day-old offspring was detected by the immunofluorescence (IF) assay. The expression of TLR4, NF-κB p65 and other related proteins in the ovarian tissue was determined by Western blot. ResultCompared with the control group, the model group showed reduced primordial follicles in the offspring on the day of birth (P<0.01) as well as disturbed estrous cycle, decreased ovary index and uterus index (P<0.01), reduced corpus luteum, increased atretic follicles (P<0.01), lowered serum levels of AMH and E2 (P<0.01), elevated serum levels of LH, FSH, IL-1β, and TNF-α (P<0.05, P<0.01), and up-regulated protein levels of TLR4, NF-κB p65, recombinant myeloid differentiation factor 88 (MyD88), and phosphorylated NF-κB inhibitor (p-IκBα) (P<0.01) in the 75-day-old offspring rats. Compared with the model group, Zuoguiwan and vitamin E increased the primordial follicles in the offspring on the day of birth (P<0.01). Moreover, they resumed the estrous cycle, increased the ovary and uterine indices (P<0.05, P<0.01) and corpus luteum (P<0.01), reduced atretic follicles (P<0.01), elevated the serum levels of AMH and E2 (P<0.05, P<0.01), lowered the serum levels of LH, FSH, IL-1β, and TNF-α (P<0.05, P<0.01), and down-regulated the expression of TLR4, NF-κB p65, MyD88, and p-IκB-α (P<0.05, P<0.01) in the 75-day-old offspring. ConclusionZuoguiwan can improve the ovarian reserve in the offspring rat model of congenital kidney deficiency by regulating the TLR4/NF-κB p65 signaling pathway.
2.Analysis of hemagglutinin and neuraminidase gene characteristics of influenza B (Victoria) viruses isolated in Xi′an city from 2019 to 2023
Yangni DENG ; Peng ZHANG ; Yujie YANG ; Yuewen HAN ; Yuanyuan DUAN ; Shuling LI ; Yinjuan HUANG ; Jianing WEI ; Ye ZHANG ; Rui WU ; Hailong CHEN
Chinese Journal of Experimental and Clinical Virology 2024;38(5):552-557
Objective:To investigate the genetic and evolutionary characteristics of hemagglutinin (HA) and neuraminidase (NA) genes of influenza B/Victoria (BV) virus in Xi′an city from 2019 to 2023.Methods:Twenty-five BV strains isolated from the Xi′an influenza surveillance network laboratory between 2019 and 2023 were collected. The HA and NA genes were sequenced using MiniSeq high-throughput sequencing platform. An evolutionary tree was constructed using bioinformatics software to analyze homology and mutation sites, and to predict N-glycosylation sites online. The antigenicity of the strains was analyzed through hemagglutination inhibition tests.Results:The BV influenza in Xi′an exhibited a distinct seasonal transmission pattern from 2019 to 2023, with peak prevalence occurring during the winter and spring seasons. The evolutionary analysis of the HA genes shows that the strains from Xi′an in 2019 belong to the V1A.3 branch, and the strains from 2021 to 2023 belong to the V1A.3a.2 branch. Analysis of antigenic sites showed that there were variations in 6 sites of 3 antigenic determinants in the HA proteins of the BV strains from 2021-2022 compared to 2019, and 2 sites of 1 antigenic determinant changed in the HA proteins in 2023 compared to 2021-2022. The evolutionary analysis of the NA genes indicates that the BV strains from Xi′an in 2019 belong to the A. 1.1 branch. By 2021 and 2022, it had evolved into the A. 1.2 clade, and by 2023, it had further evolved into the B clade and its derivatives, with no strains showing mutations associated with resistance to NA inhibitors. Antigenic analysis indicated that the majority of BV strains in Xi′an were similar to the strains included in the vaccine composition. Furthermore, glycosylation analysis showed that the potential N-glycosylation sites in the HA proteins of BV strains from 2021-2023 were reduced by one compared to those from 2019, and only a few strains from 2023 displayed alterations in the potential N-glycosylation sites of the NA proteins.Conclusions:The HA and NA genes of the BV strains from 2019 to 2023 are continuously mutating and evolving into new branches. Since 2021, V1A.3a.2 has become the dominant evolutionary branch of the HA genes, while the evolutionary branches of the NA genes from 2019 to 2023 have been continuously changing.
3.Degradation of BRD4 protein with dBET1 inhibits neurotoxic effects of α-synuclein oligomers
Yinjuan ZHENG ; Piao ZHANG ; Peiting HUANG ; Yuhu ZHANG
Chinese Journal of Geriatric Heart Brain and Vessel Diseases 2024;26(5):573-577
Objective To investigate whether degradation of bromodomain-containing protein 4(BRD4)protein with dBET1 can inhibit neuroinflammation and oxidative stress caused byα-synuclein(α-syn)oligomers.Methods After BV2 cells or SH-SY5Y cells were treated with dBET1 and α-syn oligomers,the cells were divided into Control group,α-syn group,α-syn+500 and 1000 nmol/L dBET1 groups,and dBET1 group(1000 nmol/L).Real-time quantitative fluo-rescent PCR(qPCR)was used to detect the expression of inflammatory factors and anti-inflam-matory factors,and Western blotting was employed to measure the expression of BRD4,nuclear factor 2 associated factor 2(Nrf2),phosphorylated nuclear transcription factor-κB(p-NF-κB)and phosphorylated α-synuclein(p-α-synuclein).Results The α-syn+1000 nmol/L dBET1 group had lower mRNA levels of inflammatory factors and reduced protein levels of BRD4,p-NF-κB and p-α-synuclein,but higher mRNA levels of anti-inflammatory factors and decreased Nrf2 protein level(2.02±0.14 vs 0.96±0.24,P<0.05)when compared with the α-syn group.Conclusion dBET1 can inhibit neuroinflammation and oxidative stress caused by α-synuclein through p-NF-KB and Nrf2 to a certain extent,which further provides a theoretical basis for its application in the treatment of Parkinson's disease.
4.Reference values of carotid intima-media thickness and arterial stiffness in Chinese adults based on ultrasound radio frequency signal: A nationwide, multicenter study
Changyang XING ; Xiujing XIE ; Yu WU ; Lei XU ; Xiangping GUAN ; Fan LI ; Xiaojun ZHAN ; Hengli YANG ; Jinsong LI ; Qi ZHOU ; Yuming MU ; Qing ZHOU ; Yunchuan DING ; Yingli WANG ; Xiangzhu WANG ; Yu ZHENG ; Xiaofeng SUN ; Hua LI ; Chaoxue ZHANG ; Cheng ZHAO ; Shaodong QIU ; Guozhen YAN ; Hong YANG ; Yinjuan MAO ; Weiwei ZHAN ; Chunyan MA ; Ying GU ; Wu CHEN ; Mingxing XIE ; Tianan JIANG ; Lijun YUAN
Chinese Medical Journal 2024;137(15):1802-1810
Background::Carotid intima-media thickness (IMT) and diameter, stiffness, and wave reflections, are independent and important clinical biomarkers and risk predictors for cardiovascular diseases. The purpose of the present study was to establish nationwide reference values of carotid properties for healthy Chinese adults and to explore potential clinical determinants.Methods::A total of 3053 healthy Han Chinese adults (1922 women) aged 18-79 years were enrolled at 28 collaborating tertiary centers throughout China between April 2021 and July 2022. The real-time tracking of common carotid artery walls was achieved by the radio frequency (RF) ultrasound system. The IMT, diameter, compliance coefficient, β stiffness, local pulse wave velocity (PWV), local systolic blood pressure, augmented pressure (AP), and augmentation index (AIx) were then automatically measured and reported. Data were stratified by age groups and sex. The relationships between age and carotid property parameters were analyzed by Jonckheere-Terpstra test and simple linear regressions. The major clinical determinants of carotid properties were identified by Pearson’s correlation, multiple linear regression, and analyses of covariance.Results::All the parameters of carotid properties demonstrated significantly age-related trajectories. Women showed thinner IMT, smaller carotid diameter, larger AP, and AIx than men. The β stiffness and PWV were significantly higher in men than women before forties, but the differences reversed after that. The increase rate of carotid IMT (5.5 μm/year in women and 5.8 μm/year in men) and diameter (0.03 mm/year in both men and women) were similar between men and women. For the stiffness and wave reflections, women showed significantly larger age-related variations than men as demonstrated by steeper regression slopes (all P for age by sex interaction <0.05). The blood pressures, body mass index (BMI), and triglyceride levels were identified as major clinical determinants of carotid properties with adjustment of age and sex. Conclusions::The age- and sex-specific reference values of carotid properties measured by RF ultrasound for healthy Chinese adults were established. The blood pressures, BMI, and triglyceride levels should be considered for clinical application of corresponding reference values.
5.Study on correlation between treponema pallidum specific antibodies levels of syphilis and immunoglobulin levels in active syphilis patients
Yao WANG ; Jie DING ; Qian YANG ; Yinjuan SHI ; Yuting LIANG ; Yang HE ; Sheng ZHANG
Chinese Journal of Immunology 2024;40(8):1584-1589
Objective:To investigate the correlation between the level of treponema pallidum specific antibodies(TPAb)and immunoglobulin A/E/M/G(IgA/E/M/G)and IgG subtypes in active syphilis patients,so as to provide a basis for the comprehensive di-agnosis and treatment and prognosis evaluation of clinical syphilis.Methods:Active syphilis patients admitted to the First Affiliated Hospital of Soochow University in 2023 were divided into 1∶4+and 1∶16+groups according to the syphilis toluidine red unheated se-rum test(TRUST),and healthy control group were selected for the detection of TPAb by chemiluminescence microparticle immunoas-say(CMIA).The contents of IgA/E/M/G and IgG1/2/3/4 were determined by immunoturbidimetry.Results:A total of 206 patients with active syphilis(125 males and 81 females)aged(46.05±19.58)years were included in this study.Compared with the healthy control group,IgG2 was decreased and TPAb,IgA/E/M/G and IgG1/3/4 levels were increased in the active syphilis group,with statistically significant differences in TPAb,IgE and IgG1/2/3 levels(P<0.05).Compared with the healthy control group,TRUST 1∶4+group had statistical differences in TPAb,IgE and IgG2,and TPAb was significantly correlated with IgE and IgG2/4.The levels of TPAb,IgA/E/M/G and IgG1/2/3 in TRUST 1∶16+group were statistically significant,and TPAb was significantly correlated with IgE and IgG4(P<0.05).In addition,there were significant differences in TPAb,IgG and IgG1/3 between the two positive groups(P<0.05).Conclu-sion:The levels of TPAb and Ig in active syphilis patients with different TRUST titers are significantly different.TPAb combined with Ig can be used to monitor the strength of syphilis activity,IgA/M/G and IgG1/3 may be candidate biomarkers.
6.Real-world validation of the chemotherapy-induced nausea and vomiting predictive model and its optimization for identifying high-risk Chinese patients.
Linlin ZHANG ; Lili ZENG ; Yinjuan SUN ; Jing WANG ; Cong WANG ; Chang LIU ; Ming DING ; Manman QUAN ; Zhanyu PAN ; Diansheng ZHONG
Chinese Medical Journal 2023;136(11):1370-1372
7.The correlation between γδ-T lymphocyte subset in early postnatal peripheral blood and brain injury of preterm infants
Xinyun JIN ; Xiaoli ZHANG ; Yinjuan WANG ; Jiaxin LIU ; Xiaoyang WANG ; Ru ZHANG ; Yi ZHANG ; Falin XU
Chinese Journal of Applied Clinical Pediatrics 2022;37(10):743-747
Objective:To investigate the correlation between preterm infants with brain injury and the proportion of lymphocyte subsets, especially γδ-T cells in the postnatal peripheral blood, and to determine the predictive potential of γδ-T cells in the early peripheral blood in brain injury.Methods:It was a prospective study involving 106 preterm infants with gestational age less than 34 weeks who were delivered in the Department of Neonatology, the Third Affiliated Hospital of Zhengzhou University from January 1, to June 1, 2021.Relative levels of γδ-T , CD4 + T, CD8 + T, CD3 + T and total lymphocyte subsets in peripheral blood collected within the first 24 hours after birth were measured by flow cytometry.Recruited infants were divided into brain injury group (36 cases) and non-brain injury group (70 cases) according to serial cranial ultrasound and magnetic resonance imaging(MRI) at the corrected gestational age of 36-37 weeks.Differences in general conditions and the proportion of lymphocyte subsets between groups were compared by the t-test or Chi- square test.Patients in brain injury group were further divided into intracranial hemorrhage(ICH) group(8 cases), periventricular leukomalacia (PVL) group (6 cases)and diffuse white matter damage (WMD) group(22 cases). The proportion of lymphocyte subsets among the different groups was compared by One- Way ANOVA, followed by the LSD- t test. Results:The proportion of γδ-T cells in postnatal peripheral blood of preterm infants at 24 hours after birth in brain injury group was significantly lower than that of non-brain injury group [(0.09±0.12)% vs.0.15±0.13)%, t=-2.445, P=0.016]. No significant differences in the proportion of the CD4 + and CD8 + T cell subsets were found between them.Both preterm infants in PVL group and WMD group had a significantly lower proportion of γδ-T cells at 24 hours after birth compared to that of the non-brain injury group [(0.03±0.05)%, (0.07±0.09)% and (0.15±0.13)%], respectively, ( t=-2.190, -2.659, all P<0.05). Conclusions:γδ-T cells in early postnatal peripheral blood may be involved in the development of brain injury in preterm infants and they had early predictive value for preterm infants at high risk of brain injury, especially the leukomalacia and diffuse white matter injury.
8.CBCT study on the safe location of palatal microscrew implant anchorage nail between maxillary first and second molars
LI Xinghan ; LI Jun ; MENG Yitong ; TANG Yulong ; XU Jialin ; YANG Ying ; DONG Yinjuan ; ZHANG Xiaodong
Journal of Prevention and Treatment for Stomatological Diseases 2022;30(1):39-44
Objective:
Conebeam CT (CBCT) was used to measure the palatine between the maxillary first and second molars. The proximal and distal palatal widths of the maxillary first and second molar and the palatal mucosal thickness and bone tissue thickness when microscrew implant anchorage nail were implanted at different angles provided a reference for the clinical selection of microscrew implant placement.
Methods:
The image data of 90 adult patients were selected as the research object, and the jaw bone was reconstructed by scanning. In maxillary palatine, selection of distances at 12 mm, 14 mm, 16 mm, and 18 mm from the palatal apex of maxillary first molar between the maxillary first and second molar were used as measurement, measured the proximal and distal palatal widths of maxillary first and second molar and the palatal mucosal thickness and bone tissue thickness when microscrew implant anchorage nails were implanted at 30 °, 45 °, 60 °, and 90 °. SPSS 26.0 software was used for one-way ANOVA and LSD pair comparison.
Results:
The larger the angle of the microscrew implant anchorage nail was, the smaller the proximal and distal medial widths between the maxillary first and second molar, and the difference was statistically significant (P < 0.05). Compared with the 90° direction, the proximal and distal medial widths of the microscrew implant anchorage nail were larger in the 60° direction. The greater the angle of implantation, the smaller the mucosal thickness and the greater the bone tissue thickness, and the results showed a significant difference (P < 0.001). Compared with the direction of 30° and 45°, the mucosal thickness at the direction of 60° was smaller, and the bone tissue thickness was larger. The higher the position of the microscrew implant anchorage nail, the greater the width of the proximal and distal medial, and the difference was statistically significant (P < 0.05). Compared with the positions 12 and 14 mm from the palatal tip, the proximal and distal medial widths of the microscrew implant anchorage nail were larger. The higher the implant position was, the greater the mucosal thickness and the smaller the bone tissue thickness. The results showed a significant difference (P < 0.001). Compared with the position of 18 mm from the palatal tip of the maxillary first molar, the mucosal thickness was smaller and the bone tissue thickness was larger.
Conclusion
It is most appropriate to implant microscrew implant anchorage nail at least 10 mm in length in the direction of 60° at the palatal apex 16 mm from the maxillary first molar in palatine between the first and second molar.
9.Determination of 20(S)-protopanaxadiol Concentration in Human Plasma by UPLC-MS/MS
Shenglan LIU ; Zhi TANG ; Lei CHEN ; Sufen WU ; Ling ZHOU ; Yinjuan LIAO ; Jie ZHANG
China Pharmacy 2021;32(18):2248-2253
OBJECTIVE :To est ablish the method for the determination of 20(S)-protopanaxadiol(PPD)concentration in human plasma. METHODS :Plasma samples were precipitated with acetonitrile and determined by UPLC-MS/MS ,using finandrogen as internal standard. The determination was performed on Waters ACQUITY UPLC HSS T 3 column with mobile phase consisted of 5 mmol/L ammonium bicarbonate aqueous solution-acetonitrile (gradient elution )at the flow rate of 0.4 mL/min. The column temperature was set at 40 ℃,and sample size was 10 μL. The ion source was electrospray ion source,and negative ion scanning was carried out with multiple reaction monitoring mode . The ion pairs used for quantitative analysis were m/z 459.40→ 375.20(PPD)and m/z 371.30→315.30(internal standard ). At the same time ,the method was applied to the determination of clinical samples. RESULTS :The linear range of PPD was 0.25-30.00 ng/mL(r=0.999 2),and the limit of quantitation was 0.25 ng/mL. RSDs of intra-batch and inter-batch were all lower than 10%,and relative errors (RE)were -14.61%-12.69%. Extraction method and matrix effect did not affect the quantitative determination of PPD. In ginsenoside CK 100 mg group ,ginsenoside CK 200 mg group and ginsenoside CK 300 mg group ,mean cmax of patients with rheumatoid arthritis after oral administration of corresponding drugs were 18.06,30.03,27.00 ng/mL;median tmax were 12.0,6.0,12.0 h;mean AUC 0-t were 622.52,668.15, 1 155.97 ng·h/mL. CONCLUTIONS :The method for the determination of PPD concentration in human plasma is successfully established. The method is sensitive ,accurate, kq1907011) stable,easy to operate and less plasma consumption. It can be used for the quantitative determination of clinical samples.
10.Detection of 2019-nCoV nucleic acid at different time of courses in confirmed COVID-19 cases
Zerun XUE ; Haifei YOU ; Rui WU ; Yang LUAN ; Hailong CHEN ; Quanli DU ; Yuewen HAN ; Yujie YANG ; Yangni DENG ; Yinjuan HUANG ; Hao LI ; Kaixin LI ; Leile ZHANG ; Peng ZHANG ; Chaofeng MA
Chinese Journal of Experimental and Clinical Virology 2021;35(6):728-731
Objective:To explore the positive rates of 2019-nCoV nucleic acid at different time of courses of COVID-19.Methods:Patients with confirmed COVID-19 were enrolled in this study. Nasal and throat swabs were collected from different courses of disease. Real-time RT-PCR was used to detect 2019-nCoV nucleic acid.Results:From January 23 to February 20, 2020, a total of 120 confirmed cases of COVID-19 were reported in Xi’an, and 85 cases(70.83%) were positive in first detection. The COVID-19 cases included consistently nucleic acid positive and intermittently nucleic acid positive patients. 2019-nCoV nucleic acid could be detected in incubation period, and the longest observed duration of nucleic acid positive in this study was 26 days. The positive rate of 2019-nCoV nucleic acid was up to 84.21% on the 6th day, and the positive rate decreased as time passed during the course of COVID-19. Three patients (2.86%) were tested positive for 2019-nCoV nucleic acid again in nasal and throat swabs after discharge.Conclusions:The positive rate of 2019-nCoV nucleic acid was higher in the early stage of disease. 2019-nCoV nucleic acid can be detected in incubation period, and virus shedding may last for a long period.


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