1.Correlation study of serum adiponectin and brain natriuretic peptide with left ventricular hypertrophy in patients with essential hypertension
Chinese Journal of Geriatrics 2013;32(11):1152-1154
Objective To study the correlation of serum adiponectin (ANP) and brain natriuretic peptide (BNP) levels with left ventricular hypertrophy in essential hypertension patients,and to investigate its potential mechanism.Methods 80 patients with essential hypertension in our hospital were recruited.Secondary hypertension,cardiac,brain and renal insufficiency and diabetes mellitus were excluded in the study patients.Patients were divided into 2 groups:left ventricular hypertrophy group (LVH group,n=40) and hypertension group (hypertension patients without left ventricular hypertrophy,n=40).Age-,sex-,body mass matched health subjects were selected as control group (n =40).Serum levels of ANP and BNP were determined by enzyme-linked immunosorbent assay (ELISA).Left ventricular end diastolic dimension (LVEDD),left ventricular posterior wall thickness (LVPWT) and interventricular septum thickness (IVST) were detected by ultrasonic cardiography.Left ventricular mass (LVM) and left ventricular mass index (LVMI) were calculated according to formula of Devereux.Results There were significant differences in serum levels of APN and BNP and LVMI between hypertension,LVH and control groups [(7.9±0.3) mg/L,(6.7±0.1)mg/L vs.(12.0±0.2)mg/L,(116.1±16.1)μg/Lvs.(182.4±18.5)μg/L,(104.0± ±9.6)g/m2,(153.7±21.0)g/m2 vs.(83.1±9.1)g/m2,respectively,all P<0.01].Serum APN level was negatively correlated with the severity of LVH (r=-0.789,P<0.01),while serum BNP level was positively correlated with the severity of LVH (r=0.811,P< 0.05).Conclusions Serum APN and BNP levels have correlation with hypertension and hypertension related LVH.
2.Resveratrol regulates serum lipid and antioxidant enzymes level in an atherosclerotic rabbit model
Rui SONG ; Yanyu CHEN ; Wanqiu LI ; Jianlin DOU ; Ge ZHANG ; Lin SUN
Chongqing Medicine 2014;(31):4169-4171,4174
Objective This study was designed to investigate influence of resveratrol on serum lipid and antioxidant enzyme lev‐els in atherosclerotic rabbit model ,and to explore its influence on NF‐κB and MAPKs signal pathway .Methods Rabbits were as‐signed to five groups :control (group A) ,high fat diet group (group B) ,resveratrol group (group C ,D and E) .The contents of lip‐ids level (TC ,TG ,LDL‐C ,HDL‐C) and antioxidant enzyme (GSH ,GSH‐PX ,GST ,γ‐GCS ,CAT ,SOD ,MDA) levels in the serum were measured respectively and the difference was studied .Phosphorylation levels of MAPKs cascades ,NF‐κB were measured by Western blot .Results Compared with group A ,group B had elevated levels of blood lipids ,antioxidant enzymes were on the de‐cline ,the MDA content increased ,MAPKs and the NF‐κB protein phosphorylation enhanced .C ,D ,E group can reduce levels of blood lipids ,increases HDL‐C ,improve antioxidant enzyme activity and reduce MDA content ,inhibit MAPKs ,NF‐κB protein phos‐phorylation .Conclusion Resveratrol could reduce the atherosclerotic rabbit blood lipid levels ,increase antioxidant enzyme activity , reduce the MDA level and this effect is likely to inhibit NF‐κB and MAPKs signaling pathway activation .
3.Role of GABAAα3 and GABAB receptors in ventrolateral periaqueductal gray in rats with acute pain
Chao LOU ; Guizhi WANG ; Jianfeng YU ; Wenying CHI ; Wanghua JIA ; Chunyan ZHANG ; Wanqiu SUN
The Journal of Clinical Anesthesiology 2017;33(5):488-491
Objective To investigate the role of GABAAα3and GABAB receptors in the ventrolateral periaqueductal gray in the development of paw acute pain in rats.Methods Twelve male SD rats, weighing 280~320 g, were randomly divided into two groups: normal saline group (group NS), formaldehyde-induced pain group (group F), 6 rats in each group.In group F, rats were subcutaneously injected with 2% formaldehyde 50 μl into the ventral surface of right hind paw to induce periphery inflammatory pain.In group NS, rats were subcutaneously injected with normal saline into the ventral surface of right hind paw.Mechanical threshold was assessed using von Frey hairs for every ten minutes.The rat pain behavior scores were recorded for every five minutes.The thickness of skin and skin temperature were recorded for every fifteen minutes.Results Mechanical hyperalgesia were induced in group F after formalin injection into right hind paw.Compared with group NS, rat pain behavior scores were increased significantly in group F at all time points after injection, mechanical threshold were decreased significantly in group F at 10-60 min after injection, the temperature of the skin and the skin thickness were increased significantly in group F at 15-60 min after injection (P<0.05), the levels of the expression of GABAAα3 and GABAB were significantly increased in group F (P<0.05).Conclusion GABAAα3and GABAB receptors mediates formalin-induced hyperalgesia at ventrolateral portion of the PAG (vlPAG) of rats.
4.The therapeutic effects of TCDCA on cartilage degeneration in experimental temporomandibular joint os-teoarthritis in rats
Wanqiu HUO ; Mian ZHANG ; Qian LIU ; Dongmei WANG ; Meiqing WANG
Journal of Practical Stomatology 2024;40(2):198-203
Objective:To investigate the effects of the taurochenodeoxycholic acid(TCDCA)on cartilage degeneration in tem-poromandibular joint osteoarthritis(TMJ OA).Methods:36 female SD rats aged 6 weeks were randomly divided into 3 groups:con-trol group(CON),unilateral anterior crossbite group(UAC),UAC plus TCDCA injection group(UAC+TCDCA).UAC model was es-tablished in all rats in UAC and UAC+TCDCA groups.Samples were collected at 8 and 12 weeks(control group,UAC group,UAC+TCDCA group)after set up of the experiment(n=6),and TMJ morphological examination was performed.The expression of CYP7A1,BAAT and TGR5 in the tissue and cells was examined by immunohistochimical staining.Results:(1)Compared with the CON group of the same age,the cells in the condylar cartilage were disordered,the cartilage matrix was reduced and thinner in UAC group.Compared with UAC group of the same age,cell arrangement,cell number,cartilage matrix and cartilage thickness were im-proved in UAC+TCDCA group(P<0.05).(2)Compared with the CON group of the same age,the positive cells for TCDCA-specific receptor TGR5 and the key enzymes CYP7A1 and BAAT were mainly distributed in the anterior hypertrophic layer and hypertrophic layer at each time point.The number of positive cells in the UAC group was significantly reduced compared with the CON group.Conclusion:TCDCA has obvious therapeutic effects on the degeneration in TMJ OA.
5.Microanatomical study of the scapholunate interosseous ligament with micro-CT
Yujian XU ; Yongqing XU ; Haotian LUO ; Xiaoqing HE ; Xulin ZHANG ; Wanqiu ZHAO ; Huan WU ; Libo YUAN
Chinese Journal of Microsurgery 2020;43(1):56-60
Objective:To explore the morphology and vessel distribution of the scapholunate interosseous ligament and anatomical basis for the clinical reconstruction of scapholunate interosseous ligament.Methods:From October, 2018 to December, 2018, 12 fresh wrist joint specimens were perfused with gelatin-lead oxide solution from ulnar or radial artery and scanned under micro-CT. The morphology of scapholunate interosseous ligament in neutral position and the distribution of nutrient vessels in the ligament were observed on reconstructed 3D images by Mimics. The width, length and thickness of palmar, dorsal and proximal ligaments were measured. The anatomical parameters at the entrance of nutrient vessels in the scapholunate interosseous ligament were taken and their relationship with the blood supply to the scapholunate was analyzed.Results:①For scapholunate interosseous ligament, it was found that the average length of the proximal sub-region was the longest, the length of palmar and dorsal sides was similar to each other and the widest and thinnest was in palmar side, while the thickness and width of dorsal and proximal were similar. ②There was no nutrient vessel in the proximal part of the scapholunate interosseous ligament. But there were abundant nutrient vessels in the palmar and dorsal scapholunate interosseous ligament, and there was no significant difference in blood supply to palmar and dorsal scapholunate interosseous ligament ( P>0.05). ③The palmar and dorsal medial nutrient vessels that supply to the scapholunate interosseous ligament enter the scapholunate from the attachment of ligament of scapholunate interosseous joint. Conclusion:The palmar side of the scapholunate interosseous ligament is wider and thinner than that of the other subareas, which makes it more vulnerable to injury from an anatomical point of view. There is abundant blood supply to the palmar and dorsal subareas of the scapholunate interosseous ligament and the supplying vessels anastomose inside the scapholunate bone. There is no distribution of blood vessel at the proximal part of scapholunate interosseous ligament, hence is difficult to heal. An injury of palmar and dorsal ligaments may affect the blood supply of scapholunate.
6.Detection and analysis of EBV DNA integration in NK/T cell lymphoma genome
Xin WANG ; Xudong ZHANG ; Qingjiang CHEN ; Guannan WANG ; Junxia HU ; Shaoxuan WU ; Mijing MA ; Meifeng YIN ; Wanqiu YANG ; Meng DONG ; Mengjie DING ; Mingzhi ZHANG ; Linan ZHU
Chinese Journal of Clinical Oncology 2018;45(23):1194-1200
To investigate the presence of integrated Epstein-Barr virus (EBV) DNA in the NK/T cell lymphoma (NKTCL) ge-nome and analyze the integration information in the genome of NKTCL cell lines. Methods: PCR and in situ hybridization were used to detect EBV infection in five EBV (+) NK/T samples and four EBV (-) NK/T samples provided by the biobanks of the First Affiliated Hospi-tal of Zhengzhou University. Whole-genome DNA of the samples was sequenced and subjected to bioinformatics analysis. Whole-ge-nome sequence alignment was used to identify the EBV integration sequence. BLAST analysis was used to compare EBV fasta files of the samples and EBV fasta library. CREST software was used to extract softclip reads, filter all paired reads, and enumerate their distri-bution on chromosomes. The integrated genomics viewer (IGV) was used to compare the distribution of reads in partial regions of chromosome. PCR was used to amplify the high-frequency integration region of the EBV DNA. The amplified fragments were sanger se-quenced. Results: EBV DNA and EBER expression were detected in five EBV (+) NK/T samples but not in the four EBV (-) NK/T samples. Sequencing depth, coverage depth, proportion of coverage, and proportion of alignment all met the requirements for subsequent re-search. Sequence alignment revealed that the captured sequences were viral sequences. Filtered reads were most numerous in EBV (+) NKTCL cell line SNK, YTS, and EBV (+) nasal NKTCL tissue. The reads were non-randomly enriched in chromosome 2. EBV DNA inte-gration in the 400 bp region of chr2:30234084-30234483 caused insertion or deletion in the chr2p23.1 site. Conclusions: EBV DNA is highly integrated in the chr2p23.1 site of EBV (+) NKTCL cells and may affect the expression of related genes.
7.Expression and clinical significance of PD-1/PD-Ls in EBV-positive T/NK lymphoprolif-erative disorders
Junxia HU ; Qingjiang CHEN ; Xudong ZHANG ; Wencai LI ; Guannan WANG ; Xin WANG ; Meng DONG ; Shaoxuan WU ; Mijing MA ; Meifeng YIN ; Wanqiu YANG ; Mengjie DING ; Mingzhi ZHANG ; Linan ZHU
Chinese Journal of Clinical Oncology 2018;45(24):1248-1253
Objective: To investigate the expression and clinical significance of programmed death-ligand 1 (PD-L1), programmed death-ligand 2 (PD-L2), and their receptor programmed cell death protein 1 (PD-1) in EBV-positive T/NK lymphoproliferative disease [Epstein-Barr virus-positive T/natural killer (NK)-cell lymphoproliferative disease, EBV(+)-T/NK-LPD]. Methods: The pathological paraffin-embedded tissues of 17 patients with EBV(+)-T/NK-LPD from the First Affiliated Hospital of Zhengzhou University from January 2013 to December 2017 were collected. These patients include 12 males and 5 females, aged 10-82 years old, the average age being 29 years, 4 people in gradeⅠ, 7 in gradeⅡ, 3 in gradeⅢ, and 3 people with hydroa vacciniforme-like lymphoproliferative disorders. Immunohistochemical SP method was used to detect the expression of PD-1, PD-L1, and PD-L2 in human EBV(+)-T/NK-LPD tissues. The relationship between PD-1, PD-L1, PD-L2 expression, and clinicopathological parameters, pathological grades and prognosis were analyzed by Fisher's exact probabilities and Spearman rank correlation. Result: After statistical analysis, the results showed that in 17 cases of tissue samples, there were 12 cases with positive PD-1 expression, 6 cases with positive PD-L1 expression and 5 cases with positive PD-L2 expression. There was no significant correlation between PD-1 and PD-L2 expression and prognosis (P>0.05). PD-L1 expression showed a positive correlation with prognosis (P<0.05). There was no significant correlation between the expression of PD-L1 and PD-L2 with age, sex, as well as LDH and Ki-67 levels (P>0.05). Moreover, there was no significant correlation of PD-1 and PD-L2 expression with pathological grade (r=0.141, r=-0.149, both P>0.05). However, there was a negative correlation between the PD-L1 expression and pathological grade (r=-0.563), and the correlation between the PD-L1 ex-pression and pathological grade was statistically significant (P<0.05). Conclusions: PD-1, PD-L1, and PD-L2 are abnormally expressed in the pathological tissues of EBV(+)-T/NK-LPD. Although there was no significant correlation between the expression of PD-1 and prognosis or pathological grade, it was significantly higher in EBV+T/NK-LPD. PD-1/PD-Ls associated signaling pathway is expected to be a potential new target for EBV(+)-T/NK-LPD immunotherapy.
8.Effect of the transcription factor SOX4 on the proliferation, migration and apoptosis of ovarian granulosa cells
Mengmeng Fan ; Cong Li ; Xue Liang ; Wanqiu Zhang ; Huiping Wang ; Zhimin Zhai
Acta Universitatis Medicinalis Anhui 2023;58(10):1646-1651
Objective :
To investigate the impact of SOX4 on ovarian granulosa cells,stable overexpression of SOX4 was achieved in human KGN cell line,followed by analysis of its effects on proliferation,migration and apoptosis.
Methods :
The recombinant lentiviral plasmid pLV-EF1a-GFP / Puro-SOX4 was generated through homologous recombination with linearized pLV-EF1a-GFP / Puro vector.Human ovarian granulosa cells ( KGN cell line ) were transduced with Lentiviral expression vectors.KGN cells infected with pLV-EF1a-GFP / Puro-NC were served as the LV-CON group,while those infected with pLV-EF1a-GFP / Puro-SOX4 were designated as the LV-SOX4 group.Following transfection,puromycin selection was employed to establish stable SOX4-expressing KGN cells.The expres- sion levels of SOX4 m RNA and protein in KGN cells from the LV-CON and LV-SOX4 groups were assessed using RT-qPCR and Western blot analysis.Cell proliferation was assessed using the CCK-8 assay in both LV-CON and LV-SOX4 groups.Cell migration ability was evaluated by means of a cell scratch test in these two groups.The proportion of apoptotic cells was determined via flow cytometry analysis in both LV-CON and LV-SOX4 groups.
Results:
The sequencing results of pLV-EF1a-GFP / Puro-SOX4 indicated a complete match between the inserted gene se- quence and the SOX4 mRNA sequence.The lentiviral titers were 7 × 108 TU / ml in the LV-CON group and 1 × 108 TU / ml in the LV-SOX4 group.The recombinant plasmid was successfully transfected into KGN cells with a transfection efficiency of over 90% under fluorescence inverted microscopy.The results of RT-qPCR and Western blot tests demonstrated a significant increase in the expression level of SOX4 in KGN cells of LV-SOX4 group compared to that of LV-CON group (t = 3. 10,P <0. 05 ; t = 14. 20,P <0. 05) .The CCK-8 assay results demonstrated that the LV-SOX4 group exhibited a significant increase in cell proliferation (24 h : t = 45. 92,P<0. 01 ; 72 h : t = 25. 60,P <0. 01) compared to the LV-CON group.The cell scratch assay indicated that the migratory capacity of KGN cells in the LV-SOX4 group was significantly enhanced (t = 7. 65,P <0. 01) compared to that in the LV-CON group. The LV-SOX4 group exhibited a significant reduction in apoptosis ratio (t = 25. 84,P<0. 01) compared to the LV- CON group.
Conclusion
SOX4-overexpressing KGN cell line was successfully established,and the overexpression of SOX4 facilitated proliferation and migration while inhibiting apoptosis in human ovarian granulosa cells.
9.Transcriptomic analysis of chondroprogenitors in response to flow fluid shear stress
Lingfeng XU ; Yuejiao ZHANG ; Jianchang ZHANG ; Jia YU ; Wanqiu HUO ; Jiali XU ; Meiqing WANG
Journal of Practical Stomatology 2024;40(1):36-42
Objective:To investigate the response of mandibular condylar chondroprogenitors to flow fluid shear stress(FFSS).Methods:Chondroprogenitors were in vitro cultured and stimulated with FFSS that can cause cell degeneration,and treated with sec-ond-generation high-throughput RNA sequencing.Differential gene expression was screened using DESeq2 software for gene ontology(GO)functional enrichment analysis,kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis and protein-protein interaction(PPI)network analysis.qRT-PCR was performed to validate the core genes screened by PPI.Results:A total of 1996 differentially expressed genes were obtained,mainly including inflammatory response and cell cycle related molecules.Among them,Actal,Atf3,Ccl2,116,Nfkbia,Ret and Vcaml were identified as the core genes.Conclusion:FFSS stimulation affects chondroprogenitor function by acting on inflammatory responses and cell cycle-related signaling pathways in chondroprogenitors.
10.Study on the Toxicity Mechanism of Yunaconitine-induced Arrhythmia in Rats Based on Calcium Overload
Zhisi ZHANG ; Wanqiu CHENG ; Tao JIANG ; Zhibin SHEN ; Yanfen CHEN ; Cong CHEN ; Ying SITU ; Chunping TANG
China Pharmacy 2021;32(23):2854-2858
OBJECTIVE:To study the toxicity mechanism of yunacotine-induced arrhythmia in rats. METHODS :Totally 32 rats were randomly divided by random number table method into normal control group ,yunacotine low-dose and high-dose groups (0.09,0.14 mg/kg),aconitine group (positive control ,0.88 mg/kg),with 8 rats in each group. Administration groups were given the corresponding drugs once a day ,and normal control group was given the constant volume of normal saline ,for consecutive 7 d. After last intragastric administration ,the changes of electrocardiogram (ECG) were observed. The contents of adenosine triphosphate(ATP)in myocardial tissue and Ca 2+ in myocardial cells ,the activities of Na +-K+-ATPase and Ca 2+-Mg2+-ATPase as well as the protein expression of ranolidine receptor 2(RyR2)and Ca 2+-ATPase(SERCA2)in myocardial tissue were determined. RESULTS:Compared with normal control group ,time limit of QRS wave and QTc intervals of rats were prolonged significantly in yunaconitine low-dose group (P<0.01). The content of Ca 2 + in myocardial cells , the ATP contents , the activities of Ca2+-Mg2+-ATPase and Na +-K+-ATPase as well as the protein expression of SERCA 2 in myocardial tissue were reduced significantly (P<0.05 or P<0.01). The heart rate of rats in yunaconitine high-dose group and aconitine group were increased significantly (P< 0.05 or P<0.01),and time limit of QRS wave and QTc intervals were significantly prolonged (P<0.01);the content of Ca 2+ in myocardial cells was increased significantly (P<0.01);ATP content ,the activities of Ca 2+-Mg2+-ATPase and Na +-K+-ATPase,and protein expression of RyR 2 and SERCA 2 in myocardial tissue were decreased significantly (P<0.01). CONCLUSIONS : Yunaconitine can induce arrhythmia in rats ,the mechanism of which may be associated with Ca 2+ overload that resulted from reducing the activities of Na +-K+-ATPase and Ca 2+-Mg2+-ATPase and down-regulating the expression of related calcium transporter RyR2 and SERCA 2.