1.Qualitative and Quantitative Analysis of Chemical Constituents in Liu Junzitang by UPLC-Q-TOF-MS/MS and UPLC
Qiyao JIANG ; Chenchen LIU ; Huiling CHEN ; Zhifeng HUANG ; Wei ZHAO ; Ying LIANG ; Huafeng PAN ; Yue ZHUO
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(6):169-178
ObjectiveTo establish a qualitative and quantitative analysis method for chemical constituents in Liu Junzitang(LJZT), and to clarify its material basis. MethodThe chemical constituents in LJZT were analyzed by ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS), and the resulting compounds were identified by using databases, such as MassBank, PubChem, ChemSpider, Traditional Chinese Medicine Systems Pharmacology Database and Analytical Platform(TCMSP), and by combining with relevant literature. UPLC was used to establish a quantitative method for analysis of 9 compounds in LJZT, including liquiritin, hesperidin, lobetyolin, liquiritigenin, glycyrrhizic acid, nobiletin, tangeretin, atractylenolide Ⅱ and Ⅰ. ResultBy combining the relevant literature, database and MS information, a total of 79 compounds were identified from LJZT, including 31 flavonoids, 15 terpenoids, 14 nitrogen-containing compounds, 6 phenylpropanoids, 6 organic acids and 7 other compounds. The established quantitative analytical method for the nine representative components showed good linearity within their respective linear ranges, and the precision, stability, reproducibility and recovery were in accordance with the requirements. The quantitative results showed that the contents of liquiritin, hesperidin, lobetyolin, liquiritigenin, glycyrrhizic acid, nobiletin, tangeretin, atractylenolide Ⅱ and Ⅰ in LJZT were 0.376 5, 2.602 1, 0.082 6, 0.128 1, 1.778 6, 0.015 7, 0.006 7, 0.030 4, 0.003 2 mg·g-1, respectively. ConclusionThe established method can quickly, sensitively and accurately analyze the chemical constituents in LJZT, clarify that the material basis of LJZT is mainly flavonoids, terpenoids and nitrogen-containing compounds, and simultaneously determine the contents of the 9 components, which can lay a foundation for the research on quality control, mechanism and clinical application of LJZT.
2.Long non-coding RNA MALAT1 regulates astrocyte proliferation and apoptosis and affects MAPK/ERK1,2 signaling pathway
Hui HU ; Xue WANG ; Yuhan WU ; Huafeng DONG ; Ling ZHANG ; Aijun WEI ; Fang XIE ; Yun ZHAO ; Zhaowei SUN ; Lingjia QIAN
Military Medical Sciences 2024;48(5):347-354
Objective To investigate the effect of MALAT1 expressions on cell proliferation and apoptosis in astrocytes by regulating mitogen-activated protein kinase(MAPK)/extracellular signal-regulated kinase(ERK1,2)pathway.Methods The MALAT1 gene was knocked down and over-expressed in C8-D1A cells by lentiviral and plasmid vectors,respectively.The expressions of MALAT1,cell proliferation-related markers(Ki67,MCM2,PCNA)and apoptosis-related proteins(Caspase-3,Bax,Bcl-2)were detected by quantitative real-time polymerase chain reaction(qPCR).CCK-8 assay and flow cytometry were used for cell proliferation and apoptosis in C8-D1A cells.Immunofluorescence was adopted to detect the protein expressions of Caspase-3 and Ki67.Western blotting was used to detect the protein expressions of Caspase-3,Bax,Bcl-2,ERK1/2,p-ERK1/2,p38MAPK and p-p38MAPK.Results Compared with the control group,over-expressed MALAT1 inhibited cell proliferation and induced cell apoptosis in C8-D1A cells while the knockdown of MALAT1 significantly enhanced cell proliferation and anti-apoptotic ability in C8-D1A cells.The proportion of C8-D1A cells in G0/G1-phase and G2/M-phase was higher than in the control group as evidenced by flow cytometry,but was lower in S-phase.Meanwhile,data showed that Caspase-3 was increased while p-ERK1/2 was decreased in terms of protein levels.The mRNA expressions of Ki67 and PCNA were decreased.After knockdown of MALAT1,the proportion of C8-D1A cells in S-phase was higher,but was lower in G2/M-phase.The protein expressions of Caspase-3 and Bax decreased while those of p-ERK1/2 and p-p38MAPK increased.The mRNA expressions of Ki67,MCM2 and PCNA were increased.The differences were all statistically significant(P<0.05).Conclusion MALAT1 promotes astrocyte apoptosis and inhibits proliferation by regulating the MAPK/ERK1,2 signaling pathway.
3.Clinical significance of Endoglin in prostate cancer research
Hui YANG ; Hongyi ZHANG ; Huafeng LI ; Ganggang ZHAO
International Journal of Laboratory Medicine 2024;45(15):1887-1890
Prostate cancer is one of the most common malignant tumors in male genitourinary system,and it is one of the main causes of male death in Europe and America.The incidence of prostate cancer in our coun-try is increasing,the key of treatment of prostate cancer is early diagnosis and early treatment.Endoglin also known as endothelial glycoprotein,is the most reliable marker of endothelial cell proliferation,which is over-expressed in neovascularization.Soluble Endoglin was found in the serum of tumor patients,which can be used as an auxiliary diagnosis.Inhibition of Endoglin receptor can inhibit the formation of tumor blood vessels,which provides a basis for targeted therapy.Microvessel density marked by Endoglin is of great significance in the clinical and pathological grading of solid tumors and can be used to evaluate the prognosis.This article re-views the role of Endoglin in the pathogenesis,diagnosis,treatment and prognosis of prostate cancer.
4.Exploring the Mechanism of Action of Danshen Baoxin Cha on Depressed Mice with Coronary Heart Disease Based on Network Pharmacology and NLRP3 Inflammatory Pathway
Di ZHAO ; Yi HUANG ; Wenxuan CHEN ; Guoyang ZHENG ; Kairong ZHENG ; Limei YAO ; Huafeng PAN ; Weirong LI
Traditional Chinese Drug Research & Clinical Pharmacology 2024;35(10):1552-1561
Objective To explore the mechanism of action of Danshen Baoxin Cha(DBC) on depressed mice with coronary heart disease (CHD) based on network pharmacology and NLRP3 inflammatory pathway. Methods (1) TCMSP and BATMAN-TICAM databases were used to screen the DBC active ingredients and targets. The targets of CHD with depression were screened using the OMIM and Genecards databases. The targets of DBC active ingredients and related targets of CHD with depression were imported into Venny 2.1 online platform to obtain the intersection targets,which was the potential target of DBC in the treatment of CHD with depression. Protein-protein interaction (PPI) analysis was performed on the intersection targets using the STRING platform to screen the key targets. A "drug-active ingredients-disease-targets" network was created to select the main active ingredients and core targets of DBC for the treatment of CHD with depression. Thereafter,the primary targets were examined by GO function and KEGG pathway enrichment using the Metascape database.(2)Kunming mice were split into six groups of eight mice each at random:the control group,the model group,the positive control group (metoprolol tartrate 5.14 mg·kg-1+sertraline hydrochloride 10.3 mg·kg-1),and the DBC high-,middle-,and low-dose groups (30.8,15.4 and 7.7 g·kg-1·d-1). Chronic unpredictable mild stimulation (CUMS)and subcutaneous injection of isoprenaline hydrochloride (ISO) were used to induce a mice model of CHD with depression. Mice were treated orally with the corresponding drug once a day for 18 consecutive days. Behavioral experiments involving forced swimming test,tail suspension test,and open-field test were applied to detect depression levels of mice. Histopathological alterations in hippocampus tissues were noted using HE and Nissl staining. qPCR was used to determine the mRNA expression levels of IL-6,TNF-α,NLRP3,IL-1β,IL-10,and Caspase-1 in hippocampus tissues. Results(1) Sixty-five active components in Salvia and seven active components in green tea were screened out. A total of 1042 potential targets and 2116 CHD complicated with depression-related targets were obtained. The intersection of the targets of active components and disease-related targets was performed by Venny 2.1.0 platform to obtain 299 potential targets (common targets) of DBC in the treatment of CHD with depression. The core targets including IL-1β,AKT1,TNF-α,IL-6,VEGFA,CASP3 and IL-10 were screened through PPI network analysis of potential targets. Key active ingredients including vitamin B,luteolin,salvianolic acid,tanshinone ⅡA and catechin,as well as key targets,such as PTGS2、IL-1β、IL-6、TNF-α and IL-10,were obtained by network analysis of "drugs-active ingredients-disease-targets". The potential targets were correlated with biological processes such as inflammation response,regulation of tumour necrosis factor (TNF),glucocorticoid regulation,regulation of nuclear factor kappa B(NF-κB) transcription factor,as well as major pathways including PI3K-Akt signaling pathway,TNF signaling pathway,apoptosis signaling pathway,and NF-κB signaling pathway.(2) Compared with the control group,mice in the model group showed a significant decrease in the total and center distance of the open field (P<0.01) and a significant increase in the time of forced swimming and immobility time of tail suspension test (P<0.01). The mRNA expression of IL-6,TNF-α,NLRP3,IL-1β,and Caspase-1 was significantly up-regulated(P<0.01) in the hippocampus tissues,but IL-10 mRNA expression was down-regulated (P<0.05). Compared with the model group,the total and center distance in DBC high-,middle-,and low-dose groups were significantly up-regulated(P<0.05,P<0.01),and the time of forced swimming and immobility time of tail suspension test were significantly down-regulated (P<0.01). The mRNA expression of IL-6,TNF-α,NLRP3,IL-1β and Caspase-1 of the DBC high-,middle-,and low-dose groups were significantly down-regulated(P<0.01),IL-10 mRNA expression in mice hippocampus tissue of DBC high-and middle-dose groups was up-regulated (P<0.01). Conclusion The intervention effect of DBC on depressed mice with CHD may be achieved by active ingredients including luteolin,tanshinone,salvianolic acid and catechin acting on the key targets,such as IL-6,TNF-α,IL-1β and IL-10,to regulate the NLRP3 inflammatory signaling pathway.
5.Clinical effect of fluid resuscitation guided by intra-abdominal pressure and oxygenation index for severe acute pancreatitis patients
Huafeng ZHANG ; Jia ZHAO ; Yunzhong ZHANG ; Deyi LIU ; Benling HU ; Huanlun WANG ; Jinhui LI
Chinese Critical Care Medicine 2022;34(5):525-528
Objective:To investigate the effect of the liquid resuscitation therapy strategy using intra-abdominal pressure (IAP) and oxygenation index (PaO 2/FiO 2) as the end point in patients with severe acute pancreatitis (SAP). Methods:A retrospective study was performed, including 84 patients with SAP in emergency intensive care unit of Qingzhou Hospital Affiliated to Shandong First Medical University from January 2018 to August 2021. According to the status of fluid balance at admission, all patients were divided into the positive fluid balance group (43 cases) and the negative fluid balance group (41 cases). The clinical data including gender, age, etiology, underlying disease, acute physiology and chronic health evaluation Ⅱ (APACHEⅡ) and sequential organ failure assessment (SOFA) of all patients were collected. Fluid balance, PaO 2/FiO 2, IAP, compliance rate, new mechanical ventilation rate and overall hospital stay of 1 week after admission were recorded and compared between the two groups. Results:After 72 hours of treatment, the cumulative fluid balance was (5 219.5±1 038.4) mL in the positive fluid balance group; IAP was higher than that before treatment [mmHg (1 mmHg≈0.133 kPa): 11.9±2.0 vs. 11.7±2.1], but no significant difference was found ( P > 0.05); PaO 2/FiO 2 was significantly higher than that before treatment (mmHg: 299.8±51.4 vs. 220.5±50.4, P < 0.05). After 72 hours of treatment, the cumulative fluid balance in negative fluid balance group was (-3 542.4±1 310.6) mL; IAP was significantly lower than before treatment (mmHg: 11.4±1.8 vs. 15.2±1.9, P < 0.05); PaO 2/FiO 2 was significantly higher than that before treatment (mmHg: 309.9±50.9 vs. 215.4±49.7, P < 0.05). In the fluid resuscitation goals, after 72 hours of treatment, the compliance rate in the negative fluid balance group was significantly higher than that in the positive fluid balance group [82.93% (34/41) vs. 62.79% (27/43), P < 0.05]; 1 week after admission, the new mechanical ventilation rate in the negative fluid balance group was significantly lower than that in the positive fluid balance group [21.95% (9/41) vs. 41.86% (18/43), P < 0.05]; however, there was no significant difference in overall hospital stay between the two groups (days: 41.2±10.9 vs. 39.1±11.5, P > 0.05). After treatment, 70 patients survived and 14 patients died (including 9 cases in the positive fluid balance group and 5 cases in the negative fluid balance group). Conclusions:Using IAP and PaO 2/FiO 2 to guide liquid therapy could result in effective fluid resuscitation in SAP. The treatment strategy effectively improved prognosis of patients with SAP.
6.Application effect of the new model of "5G cloud plus medicine" network and linkage in treatment of patients with severe trauma
Huafeng ZHANG ; Jia ZHAO ; Yunzhong ZHANG ; Deyi LIU ; Benling HU ; Huanlun WANG ; Jinhui LI ; Xiaokai LI
Chinese Journal of Trauma 2022;38(4):359-364
Objective:To explore the effect of the new model of "5G cloud plus medicine" network and linkage in improving the therapeutic effect for patients with severe trauma.Methods:A retrospective cohort study was conducted to analyze the clinical data of 410 patients with severe trauma admitted to Qingzhou People′s Hospital affiliated to Shandong First Medical University from November 2016 to November 2020. There were 258 males and 152 females, aged 16-80 years [(45.7±16.1)years]. The injury severity score (ISS) ranged from 17 to 55 points [(28.1±7.6)points]. A total of 210 patients with severe trauma were rescued by using the new model of "5G cloud plus medicine" network and linkage from November 1, 2018 to November 30, 2020 (observation group), and another 200 patients with severe trauma were rescued by the traditional treatment mode from November 1, 2016 to October 31, 2018 were selected as the control group. Time to start rescue (time from admission to the start of rescue), CT examination time (time from consultation to completion of CT scan), time to receive blood transfusion (time from blood transfusion request to execution), residence time in emergency room, ISS at postoperative 28 days, proportion of patients with blood transfusion, success rate of rescue and mortality rate were compared between the two groups.Results:Time to start rescue [(2.4±1.1)minutes], CT examination time [(29.1±10.3)minutes], time to receive blood transfusion [(28.1±10.2)minutes] and residence time in emergency room [(3.0±1.1)hours] in observation group were significantly shorter than those in control group [(5.5±1.2)minutes, (42.8±10.1)minutes, (48.5±13.1)minutes, (5.0±1.4)hours] (all P<0.05 or 0.01). ISS was (18.7±2.8)points in observation group, significantly lower than (22.1±3.4)points in control group ( P<0.05). Proportion of patients with blood transfusion was 49.5% (104/210) in observation group, similar with 42.5% (85/200) in control group ( P>0.05). Success rate of rescue was 99.0% (208/210) in observation group, significantly higher than 93.0% (186/200) in control group ( P<0.05). The mortality rate was 4.3% (9/200) in observation group, significantly lower than 8.5% (17/200) in control group ( P<0.05). Conclusion:For patients with severe trauma, the new model of "5G cloud plus medicine" network and linkage can effectively shorten the time to start rescue, CT examination time, time to receive blood transfusion and residence time in emergency room, improve the success rate of rescue and reduce the mortality rate, which is worthy of further promotion.
7.A case report of large cell undifferentiated carcinoma of the urinary bladder
Huafeng ZHANG ; Jia ZHAO ; Long XIA ; Shangdong YANG ; Xuezhong CHEN
Chinese Journal of Urology 2020;41(2):149-150
The clinical pathological data of a patient large cell undifferentiated bladder carcinoma was retrospectively analyzed and understand.The clinical and imaging findings of large cell undifferentiated bladder carcinoma was nonspecific.Diagnosis depended on the pathological and immuno-histochemical staining.The tumor is aggressive with high risk of recurrence.It is mainly treated with radical resection.
8.Analysis of prenatal phenotype and pathogenetic variant in a fetus with Papillorenal syndrome.
Xiang ZHAO ; Dan YANG ; Yumin JIA ; Yanling SHOU ; Liming WANG ; Xiangzhi WANG ; Jiena FU ; Huafeng GUO ; Jianping ZHAO ; Hao YIN ; Xueyan ZHANG ; Xiwei ZHU ; Lijuan GAO ; Chaojie MA ; Zedan XIE ; Man SHI
Chinese Journal of Medical Genetics 2020;37(9):958-961
OBJECTIVE:
To determine the carrier rate of deafness-related genetic variants among 53 873 newborns from Zhengzhou.
METHODS:
Heel blood samples of the newborns were collected with informed consent from the parents, and 15 loci of 4 genes related to congenital deafness were detected by microarray.
RESULTS:
In total 2770 newborns were found to carry deafness-related variants, with a carrier rate of 5.142%. 1325 newborns (2.459%) were found to carry heterozygous variants of the GJB2 gene, 1071 (1.988%) were found with SLC26A4 gene variants, 205 were found with GJB3 gene variants (0.381%), and 120 were found with 12S rRNA variants (0.223%). Five newborns have carried homozygous GJB2 variants, two have carried homozygous SLC26A4 variants, five have carried compound heterozygous GJB2 variants, and four have carried compound heterozygous SLC26A4 variants. 33 neonates have carried heterozygous variants of two genes at the same time.
CONCLUSION
The carrier rate of deafness-related variants in Zhengzhou, in a declining order, is for GJB2, SLC26A4, GJB3 and 12S rRNA. The common variants included GJB2 235delC and SLC26A4 IVS7-2A>G, which are similar to other regions in China. To carry out genetic screening of neonatal deafness can help to identify congenital, delayed and drug-induced deafness, and initiate treatment and follow-up as early as possible.
9. Prognostic value of pretreatment albumin to globulin ratio in prostate cancer patients treated with maximal androgen blockade
Huafeng ZHANG ; Jia ZHAO ; Daoming XU ; Long XIA
Chinese Journal of Geriatrics 2019;38(10):1153-1157
Objective:
To investigate the prognostic value of pretreatment albumin to globulin ratio(AGR)in prostate cancer(Pca)patients treated with maximal androgen blockade(MAB).
Methods:
Clinical and pathological data of 210 Pca patients who underwent MAB as first-line therapy between January 2013 and June 2018 were retrospectively analyzed.The ages of patients in our cohort ranged from 61 to 90 years, with a mean of(77.0±6.5)years.According to the cut-off point for AGR calculated by the receiver-operating curve analysis, patients were categorized into two groups: the high-AGR group and the low-AGR group.Clinical and pathological features were compared between the groups.Independent factors affecting prognosis were analyzed by using univariate and multivariate analysis.
Results:
The median follow-up duration was 44.0 months.Of the 210 patients, 99 cases had castration resistance, 100 patients(47.6%)had disease progression and 67 patients(31.8%)died.The cut-off point for AGR calculated by the receiver-operating curve analysis was 1.56.There were 103 cases in the low-AGR group(AGR<1.56)and 107 cases in the high-AGR group(AGR≥1.56). Univariate analysis revealed that the progression-free survival(PFS), cancer-specific survival(CSS)and overall survival(OS)were lower in the low-AGR group than in the high-AGR group[1.773(1.298~2.442), 1.948(1.220~3.213), 1.965(1.217~2.996), all
10.MICM classification for a case of acute myelomonocytic leukemia M
Liping HOU ; Yongping PENG ; Xiaoli SU ; Miaojuan FENG ; Ziwei CHANG ; Shan GAO ; Shuchun YU ; Zhiqiang TIAN ; Yan YU ; Xiaoli XIN ; Wenjing CUI ; Yaping ZHAO ; Guangxun GAO ; Huafeng ZHU
Chinese Journal of Clinical Laboratory Science 2019;37(6):497-471
Objective:
To describe the MICM (morphology, immunology, cytogenetics and molecular biology) characteristics of a case of acute myelomonocytic leukemia M 4C .
Methods:
The medical history data of the case of M 4C admitted to our hospital was reviewed. The results of bone marrow cell morphology, cytochemical stains, bone marrow biopsy, immunophenotype, cytogenetics, molecular test and NGS (next-generation sequencing) of the case were analyzed.
Results:
The bone marrow smear showed markedly active proliferation of bone marrow cells in which the myelomonocytic cells accounted for 85.6%. Cytochemical stains showed peroxidase (POX) stain partially and weakly positive; specific esterase AS-DCE partially positive; non-specific esterase α-NBE partially positive and smothered by sodium fluoride; non-specific esterase AS-DAE partially positive and smothered by sodium fluoride. Bone marrow biopsy showed hyperproliferative cells and diffused hyperplasia of blasts. Immunophenotype analysis showed that the abnormal cell population was positive for CD11B, CD64, CD56, cMPO, CD33, CD41, CD61, CD38 and CD58, but negative for CD13, CD34, CD117, CD7, CD123, HLA-DR, CD10, CD19, CD20, CD2, CD14, CD235, CD15, CD303, CD304, CD25, cCD79a, cCD3, cCD22, CD1a and TDT. Cytogenetic analysis showed 47, XY, t(9;11) (p22;q23),+mar. The molecular test for leukemia showed MLLT3/KMT2A gene rearrangement. NGS showed NRAS and TET2 mutation. The case was finally diagnosed as AML (acute myelomonocytic leukemia) M 4C with t(9;11)(p22;q23), MLLT3-KMT2A.
Conclusion
Leukemia M 4C may show the characteristics of both granulocytes and monocytes with complex morphological features. The combined examination of MICM should be necessary for the diagnosis of M 4C with great significance.

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