1.Cyclocarya paliurus Polysaccharide Inhibits Benign Prostatic Hyperplasia by Reducing 5α-Reductase 2
Qinhui DAI ; Mengxia YAN ; Chen WANG ; Chenjun SHEN ; Chenying JIANG ; Bo YANG ; Huajun ZHAO ; Zhihui ZHU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):107-114
ObjectiveTo investigate the effect and mechanism of polysaccharide in water extract of Cyclocarya paliurus (CPWP) in inhibiting benign prostatic hyperplasia (BPH). MethodsCPWP was obtained by heating reflux, aqueous extraction, alcohol precipitation, and freeze drying. The chemical composition and structural properties of CPWP were analyzed by high performance liquid chromatography with 1-pheny-3-methyl-5-pyrazolone pre-column derivatization and infrared spectroscopy. Male SD rats were randomly assigned into control, model, finasteride (ig 5 mg·kg-1), and low-, medium-, and high-dose (ig 50, 75, 100 mg·kg-1) CPWP groups, with 8 rats in each group. The BPH model was established by subcutaneously injecting propionate testosterone in castrated rats. The rats in the drug intervention groups were administrated with corresponding drugs, and those in the control group were administrated with an equal volume of normal saline each day. After 30 consecutive days, the rats were sacrificed, and the prostate tissue was separated and weighed. The effects of drug interventions on the body weight, prostate wet weight, and prostate index of rats were examined. The prostate tissue was stained with hematoxylin-eosin (HE) for observation of pathological changes. Enzyme-linked immunosorbent assay was employed to measure the level of dihydrotestosterone (DHT), and immunohistochemical staining was used to detect the expression of steroid 5 alpha-reductase 2 (SRD5A2) and Ki67 in the prostate tissue. ResultsCPWP was identified as a saccharide, with characteristic absorption peaks of saccharides. CPWP showed the total sugar content of 44.15% and molecular weight within the range of 5.5-78.8 kDa, being composed of mannose, rhamnose, galacturonic acid, glucose, galactose, xylose, and arabinose. Compared with the control group, the model group had significantly increased prostate wet weight and prostate index (P<0.01), thick and tall prostate epithelial cells, increased internal wrinkles, papillary expansion into the cavity, an elevation in DHT level in the serum, and up-regulated expression of SRD5A2 and Ki67 in the prostate tissue (P<0.05, P<0.01). Compared with the model group, both the finasteride and CPWP groups showed decreases in prostate wet weight and prostate index (P<0.05, P<0.01), thinned prostate epithelial cells, with only a small portion of internal wrinkles and papillary expansion into the cavity, shortened papillary protrusions, lowered DHT level in the serum, and down-regulated expression of SRD5A2 and Ki67 in the prostate tissue (P<0.01). Moreover, CPWP exerted effects in a dose-dependent manner. ConclusionCPWP inhibits BPH by regulating the expression of SRD5A2.
2.Bioinformatics analysis and validation of the interaction between PML protein and TAB1 protein
Jiacong CHENG ; Zhihui LI ; Yao LIU ; Cheng LI ; Xin HUANG ; Yinxin TIAN ; Fubing SHEN
Journal of Southern Medical University 2024;44(1):179-186
Objective To analyze the interaction between PML protein and TAB1 protein using bioinformatic approaches and experimentally verify the results.Methods Using Rosetta software,a 3D model of TAB1 protein was constructed through a comparative modeling approach;the secondary structure of PML protein was retrieved in the PDB database and its crystal structure and 3D structure were resolved.Zdock 3.0.2 software was used to perform protein-protein docking of PML and TAB1,and the best conformation was extracted for molecular structure analysis of the docking model.The interaction between the two proteins was detected using immunoprecipitation in α-MMC-treated M1 inflammatory macrophages.Results When 6IMQ of PML was used as the docking site,PML protein formed 3 salt bridges,6 hydrogen bonds and 6 hydrophobic interactions with TAB1 proteins;when 5YUF of PML was used as the docking site,PML protein formed 1 hydrogen bond,3 electrostatic interactions and 9 hydrophobic interactions with TAB1 proteins,and both of the docking modes formed good molecular docking and interactions.In the M1 inflammatory macrophages treated with α-MMC for 4 h,positive protein bands of PML and TAB1 were detected in the cell lysates in PML-IP group.Conclusion PML protein can interact strongly with TAB1 protein.
3.Association between baseline hemoglobin level and early neurological deterioration after intravenous thrombolysis in patients with acute ischemic stroke
Tinghuan WANG ; Wansi ZHONG ; Zhicai CHEN ; Ke SHEN ; Huiya YE ; Zhihui YU ; Jia LUO ; Jun MA ; Min LOU
Journal of Zhejiang University. Medical sciences 2024;53(2):168-174
Objective:To investigate the association between baseline hemoglobin level and early neurologic deterioration(END)after intravenous thrombolysis in patients with acute ischemic stroke(AIS).Methods:Data of AIS patients who received intravenous thrombolytic therapy at multiple hospitals across the country between January 2017 and July 2020 were collected from the online database Acute Stroke Patients for Stroke Management Quality Evaluation(CASE-Ⅱ,NCT04487340).Binary logistic regression analysis was used to study the factors affecting the occurrence of END after intravenous thrombolytic therapy,and the correlation between baseline hemoglobin level and END was investigated by limiting cubic spline curve analysis.Results:A total of 8162 patients were included.Patients with END had lower baseline hemoglobin levels(136 and 140 g/L,P<0.01)and higher rates of anemia(24.2%and 16.9%,P<0.01)compared with non-END patients.Binary logistic regression analysis showed that baseline hemoglobin level(OR=0.995,95%CI:0.991-0.999,P<0.05)and anemia(OR=1.238,95%CI:1.055-1.454,P<0.01)were independently correlated with the occurrence of END after intravenous thrombolysis in AIS patients.Restricted cubic spline regression showed that there was a U-shaped relationship between hemoglobin level and the risk of END after intravenous thrombolysis in AIS patients(P<0.01),although this relationship was only significant in male patients(P<0.05)and not in female patients(P>0.05).Conclusion:There is a correlation between baseline hemoglobin level and the risk of END in AIS patients after intravenous thrombolysis,especially in male patients,in whom both lower and higher hemoglobin level may increase the risk of END.
4.Bioinformatics analysis and validation of the interaction between PML protein and TAB1 protein
Jiacong CHENG ; Zhihui LI ; Yao LIU ; Cheng LI ; Xin HUANG ; Yinxin TIAN ; Fubing SHEN
Journal of Southern Medical University 2024;44(1):179-186
Objective To analyze the interaction between PML protein and TAB1 protein using bioinformatic approaches and experimentally verify the results.Methods Using Rosetta software,a 3D model of TAB1 protein was constructed through a comparative modeling approach;the secondary structure of PML protein was retrieved in the PDB database and its crystal structure and 3D structure were resolved.Zdock 3.0.2 software was used to perform protein-protein docking of PML and TAB1,and the best conformation was extracted for molecular structure analysis of the docking model.The interaction between the two proteins was detected using immunoprecipitation in α-MMC-treated M1 inflammatory macrophages.Results When 6IMQ of PML was used as the docking site,PML protein formed 3 salt bridges,6 hydrogen bonds and 6 hydrophobic interactions with TAB1 proteins;when 5YUF of PML was used as the docking site,PML protein formed 1 hydrogen bond,3 electrostatic interactions and 9 hydrophobic interactions with TAB1 proteins,and both of the docking modes formed good molecular docking and interactions.In the M1 inflammatory macrophages treated with α-MMC for 4 h,positive protein bands of PML and TAB1 were detected in the cell lysates in PML-IP group.Conclusion PML protein can interact strongly with TAB1 protein.
5.Analysis of clinical significance of serum S100A4 and S100A12 in patients with severe craniocerebral injury
Limei YANG ; Hao GUO ; Liping YANG ; Taorui SHEN ; Zhihui DONG ; Zhiyuan SONG
International Journal of Laboratory Medicine 2024;45(11):1358-1362,1367
Objective To investigate the clinical significance of serum S100 calcium binding protein(S100 A4)and S100A12 in patients with severe traumatic brain injury(sTBI).Methods A total of 147 sTBI pa-tients admitted to Handan Central Hospital from March 2021 to March 2023 were selected as the sTBI group,and 50 healthy subjects who underwent physical examination in Handan Central Hospital during the same pe-riod were selected as the control group.The correlation between S100A4,S100A12 levels and brain injury markers and the influencing factors of early death in sTBI patients were analyzed,and the predictive value of serum S100A4 and S100A12 for early death in sTBI patients was studied.Results The serum levels of S100A4,S100A12,myelin basic protein(MBP),S100B and neuron specific enolase(NSE)in sTBI group were significantly higher than those in control group(P<0.05).The serum levels of S100A4 and S100A12 were positively correlated with MBP,NSE and S100B in sTBI patients(P<0.05).Multivariate Logistic regression analysis showed that decreased Glasgow coma scale(GCS)score on admission and increased serum levels of S100A4,S100A12,MBP,NSE and S100B were risk factors for early death in sTBI patients(P<0.05).Receiv-er operating characteristic curve showed that the combination of serum S100A4 and S100A12 with GCS score,MBP,NSE and S100B was superior to any single detection in predicting early death in sTBI patients.Conclu-sion The serum levels of S100A4 and S100A12 are increased in sTBI patients,which are related to the aggra-vations of brain injury and early death.The combined detection of S100A4 and S100A12 has a good predictive value for early death in sTBI patients.
6.Epidemiological characteristics of Mycoplasma pneumoniae infection and diagnostic value of IL-6 and PCT detection in 1 122 pneumonia patients in Nanjing,China
Anran XIE ; Zhihui DONG ; Hongpan XU ; Han SHEN
Chinese Journal of Clinical Laboratory Science 2024;42(8):624-627
Objective To analyze the characteristics and epidemic trends of Mycoplasma pneumoniae(MP)infection among the pa-tients in our hospital,and explore the diagnostic values of interleukin-6(IL-6)and procalcitonin(PCT)detection for MP infection.Methods A retrospective analysis was conducted on the test results of MP nucleic acid and clinical data,including age,gender,sam-pling date,and treatment category of the 1 122 pneumonia patients in our hospital from September 1,2022,to August 31,2023.The positive detection rate of MP nucleic acid was analyzed in different genders,ages,visit categories,and seasons.Receiver operating characteristic(ROC)curves were plotted to analyze the diagnostic value of IL-6 and PCT levels,and their ratio values in Mycoplasma pneumoniae pneumonia(MPP).Results Among the 1 122 pneumonia patients,the overall positive detection rate of MP was 24.96%.The positive detection rates of MP nucleic acid in male and female patients were 23.28%and 27.29%,respectively,with no statistical-ly significant difference(P>0.05).The median age of MP nucleic acid-negative patients was 37 years,while that of MP nucleic acid-positive patients was 7 years,showing a statistically significant difference(P<0.01).MP nucleic acid-positive patients were mainly concentrated in the age groups of 1 to 2 years(20.37%),3 to 5 years(25.60%),and 6 to 8 years(67.40%).The positive detection rates of MP nucleic acid among 913 inpatients and 209 outpatients were 22.45%and 35.89%,respectively,with statistically significant difference(P<0.01).The highest MP nucleic acid positive detection rate was observed in autumn(42.47%).MP could be co-infected with five other respiratory pathogens in which human rhinovirus(HRV)was the most common(17 cases).ROC curve analysis showed that the IL-6/PCT ratio had the best diagnostic performance for MP infection in the patients with all ages,with an area under the curve(AUCROC)of 0.746,superior to IL-6 and PCT alone.For MP-susceptible patients aged 6 to 8 years,IL-6 had the highest predictive value for MP infection,with an AUCROC of 0.767.Conclusions MPP tends to be epidemic in autumn,with children aged 6 to 8 years being the main susceptible population.The IL-6/PCT ratio is expected to become an auxiliary diagnostic and prognostic evaluation indi-cator for MPP.IL-6 levels have the highest predictive value for MP infection in the susceptible population.
7.GPR120 mediates mechanism of protective effect of NF-κB and MAPK in regula-ting LTA-induced Mac-T cells
Siqi WANG ; Peiyao ZHOU ; Quanzhou MOU ; Lin WAN ; Xinli LI ; Yang LI ; Xingli HE ; Zhaoyuan WANG ; Zi WANG ; Ziqiang GAO ; Zhihui ZHAO ; Binglei SHEN
Chinese Journal of Veterinary Science 2024;44(10):2165-2171
Lipophosphatidic acid(LTA)was used to stimulate Mac-T cells,and the expression lev-els and phosphorylation levels of key proteins of nuclear factor-κB(NF-κB)and mitogen-activated protein kinase(MAPK)signaling pathway and the expression levels of upstream key action factors TLR4 and MyD88 proteins were detected by Western blot,and EDU assay was used to detect cell proliferation levels and flow cytometry was used to detect apoptosis.The results showed that acti-vation of GPR120 significantly decreased the phosphorylation levels of LTA-induced NF-κB(P65 and IκBα)(P<0.01)and MAPK(JNK,ERK,p38)(P<0.01)in Mac-T cells;inhibition of GPR120 was able to upregulate LTA-induced NF-κB(p65 and IκBα)in Mac-T cells(P<0.01)and MAPK(JNK,ERK,p38)phosphorylation levels(P<0.01);and activation of GPR120 significantly allevia-ted LTA-induced upregulation of TLR4 and MyD88(P<0.01);inhibition of GPR120 significantly exacerbated LTA-induced upregulation of TLR4 and MyD88(P<0.05);LTA stimulation led to a trend of diminished Mac-T cell proliferation and significantly increased apoptosis,whereas activa-tion of the GPR120 gene significantly increased cell activity(P<0.01),promoted cell proliferation and significantly reduced apoptosis(P<0.05)thereby alleviating the damage to Mac-T cells by LTA;LTA stimulation led to a highly significant increase in apoptosis(P<0.01).In contrast,acti-vation of the GPR120 gene significantly reversed the increase in the apoptosis rate of Mac-T cells induced by LTA(P<0.01),while inhibition of the GPR120 gene enhanced the apoptosis-promo-ting effect of LTA(P<0.05),indicating that activation of the GPR120 gene attenuated the in-crease of apoptosis rate caused by LTA-induced inflammatory Mac-T cells.The results suggest that GPR120 can regulate inflammation by mediating TLR4 and MyD88 expression to inhibit NF-κB/MAPK inflammatory pathway activation and can promote cell proliferation.
8.Clinical significance of lympho-vascular space invasion in different molecular subtypes of endometrial carcinoma
Yimeng LI ; Zhuoyu ZHAI ; He LI ; Liwei LI ; Zhihui SHEN ; Xiaobo ZHANG ; Zhiqi WANG ; Jianliu WANG
Chinese Journal of Obstetrics and Gynecology 2024;59(8):617-627
Objective:To analyze the lympho-vascular space invasion (LVSI) in different molecular subtypes of the cancer genome atlas (TCGA) molecular subtypes of endometrial cancer (EC) and to evaluate the prognostic value of LVSI in EC patients with different molecular subtypes.Methods:A total of 258 patients diagnosed EC undergoing surgery in Peking University People′s Hospital from January 2016 to June 2022 were analyzed retrospectively. Among 258 patients, 14 cases were classified as POLE-ultramutated subtype, 43 as high-microsatellite instability (MSI-H) subtype, 155 as copy-number low (CNL) subtype, and 46 as copy-number high (CNH) subtype. Fifty-four patients were positive for LVSI, while 203 tested negative.Results:(1) The incidence of LVSI was found to be highest in the CNH subtype (32.6%,15/46), followed by the MSI-H subtype (27.9%, 12/43), the CNL subtype (16.9%, 26/154), and the POLE-ultramutated subtype (1/14), with statistically significant differences ( χ2=7.79, P=0.044). (2) Staging and deep myometrial invasion were higher in the LVSI positive group than those in the LVSI negative group (all P<0.05), except for the POLE-ultramutated subtype. The grade, lymph node metastasis, and the expression of nuclear antigen associated with cell proliferation (Ki-67) were significantly higher in LVSI positive patients than those in LVSI negative EC patients with both MSI-H and CNL subtypes (all P<0.05). In CNL subtypes patients, LVSI was also associated with age, histology subtype,and progesterone receptor (PR; all P<0.05). (3) Of the 257 EC patients, 25 cases recurred during the follow-up period, with a recurrence rate of 9.7% (25/257); among them, the recurrence rate of LVSI positive patients was 22.2% (12/54), which was significantly higher than those with LVSI negative (6.4%, 13/203; χ2=12.15, P<0.001). During the follow-up period, none of the 14 patients with POLE-ultramutated had recurrence; among CNL patients, the recurrence rate was 19.2% (5/26) in LVSI positive patients, which was significantly higher than that in LVSI negative ones (5.5%, 7/128; χ2=3.94, P=0.047); where as no difference were found in both MSI-H [recurrence rates in LVSI positive and negative patients were 2/12 and 9.7% (3/31), respectively] and CNH subtype [recurrence rates between LVSI positive and negative patients were 5/15 and 9.7% (3/31), respectively] EC patients (both P>0.05). After log-rank test, the 3-year recurrence free survival (RFS) rate were significantly lower in LVSI positive patients from CNL subtype and CNH subtype than those in LVSI negative patients (CNL: 80.8% vs 94.5%; CNH: 66.7% vs 90.3%; both P<0.05). (4) Lymph node metastasis ( HR=6.93, 95% CI: 1.15-41.65; P=0.034) had a significant effect on the 3-year RFS rate of EC patients with MSI-H subtype. Multivariate analysis revealed that PR expression ( HR=0.04, 95% CI: 0.01-0.14; P<0.001) was significantly associated with the 3-year RFS rate of CNL subtype patients. Conclusions:LVSI has the highest positivity rate in CNH subtype, followed by MSI-H subtype, CNL subtype, and the lowest positivity rate in POLE-ultramutated subtype. LVSI is significantly associated with poor prognosis in CNL subtype patients and may affect the prognosis of CNH subtype patients. However, LVSI is not an independent risk factor for recurrence across all four TCGA molecular subtypes.
9.Genome characteristics of influenza B virus in Suzhou city, 2021-2022
Zefeng DONG ; Ruimin YANG ; Yang LIU ; Di WANG ; Zhihui XU ; Xuan YUAN ; Qiang SHEN ; Yuanyuan PANG ; Yu XIA
Chinese Journal of Experimental and Clinical Virology 2023;37(2):178-183
Objective:To understand the genome and genetic evolution characteristics of influenza B virus (FluB) in Suzhou city from July 2021 to January 2022.Methods:Real-time fluorescence reverse transcription polymerase chain reaction (Real-Time RT-PCR) was used for the typing of influenza virus (Flu). The detected FluB strains were sequenced by Miseq high-throughput sequencing platform through whole genome capture and library construction. The FluB hemagglutinin (HA), neuraminidase (NA) and matrix protein (MP) gene phylogenetic trees were constructed by Neighbor-Joining method (NJ) with MEGA X software. The Potential N-glycosylation sites of HA and NA proteins were predicted by NetNGlyc 1.0 server software.Results:FluB was detected in 280 of the 1 500 throat swab samples, and the FluB genome sequence was completed in 53 strains. The nucleic acid identity of 8 gene fragments (PB1, PB2, PA, HA, NP, NA, MP, NS) in the FluB strains was 99.3%-100%, 98.1%-100%, 98.8%-100%, 98.0%-100%, 99.2%-100%, 98.4%-100%, 98.2%-100% and 99.0%-100%, respectively. Except for the 4 samples in July 2021, which belonged to the V1A.3 clade of FluB, the rest of the samples belonged to the V1A.3a.2 clade. Every amino acid sequence of HA protein of Flu B collected after October 2021 showed 9-11 substitutions compared with the reference strain (B/Washington/02/2019), which sharing 9 mutation sites (H122Q, A127T, R133G, P144L, N150K, G184E, N197D, K203R and R279K). No drug-resistant mutations associated with NA inhibitors such as oseltamivir were found. Respectively, 11 and 4 potential glycosylation sites were identified in HA and NA proteins of the FluB strains.Conclusions:From July 2021 to January 2022, V1A.3a2 was the dominant FluB strains in Suzhou city, and the amino acid sequences of HA and NA proteins showed multiple site mutations.
10.A biomechanical analysis of cannulated screws fixation in a configuration of "axial compression and lateral buttress" in treatment of Pauwels type Ⅱ femoral neck fractures
Zhihui ZHONG ; Yanbin LIN ; Yan ZHUANG ; Zhaoqing SHEN ; Zhitao SU ; Fuyi ZHUANG ; Xiaoxia CHEN ; Chunling WU
Chinese Journal of Orthopaedic Trauma 2022;24(3):253-257
Objective:To investigate the biomechanical properties of cannulated screws fixation in a configuration of "axial compression and lateral buttress" in the treatment of Pauwels type Ⅱ femoral neck fractures.Methods:Ten specimens of Sawbones artificial femur were first made into models of type Ⅱ femoral neck fracture with a Pauwells angle of 50° and then randomized into 2 equal groups ( n=5). The specimens in the experimental group were subjected to fixation with cannulated screws in a configuration of "axial compression and lateral buttress" in which the axial screw was 8.5 mm in diameter and the lateral screw 6.5 mm in diameter. The specimens in the control group were subjected to conventional fixation with cannulated screws in a configuration of "inverted triangle and parallel compression" in which the 3 screws was 7.3 mm in diameter. Finally, the specimens were placed onto a biomechanical testing machine to determine the parameters of static axial stiffness, displacement under 60 to 600 N load for 5,000 cycles, ultimate load and ultimate stiffness in turn. The 2 groups were compared to find out their differences. Results:The static axial stiffness was (1,492.00 ± 87.86) N/mm, significantly higher than that in the control group [(1,200.22 ± 228.06) N/mm] ( P<0.05). There was no significant difference between the 2 groups in the cyclic load displacement [(0.44 ± 0.01) mm versus (0.57 ± 0.17) mm] ( P>0.05), but the experimental group showed a lower trend. The ultimate load and ultimate stiffness were (4,292.61 ± 804.29) N and (1,623.55 ± 180.94) N/mm in the experimental group and (4,383.64 ± 1,423.24) N and (1,433.77 ± 289.93) N/mm in the control group, showing no significant difference between the 2 groups ( P>0.05). Conclusion:In the treatment of Pauwels type Ⅱ femoral neck fractures, fixation with cannulated screws in a configuration of "axial compression and lateral buttress" may exhibit better biomechanical properties than that in a conventional configuration of "inverted triangle" .

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