1.Nutlin-3a regulates mouse adipose function by inhibition of CIDEC expression
Enhui CHEN ; Jiahui YANG ; Wei ZHAO ; Xianghong XIE ; Yanfang GUO ; Xiaojun LIU ; Li YAN
Basic & Clinical Medicine 2024;44(2):154-158
Objective To investigate the effect of Nutlin-3a,a mouse double minute 2 homolog(MDM2)inhibitor,on lipid metabolism of mouse adipose.Methods High-fat diet-induced obesity(DIO)C57BL/6J mice were randomly divided into a control group injected with DMSO and an experimental group injected with Nutlin-3a.Then we conducted glucose tolerance(GTT)and insulin tolerance(ITT)tests.The epididymal white adipose tissue(eWAT),inguinal white adipose tissue(iWAT)and brown adipose tissue(BAT)of animals were isolated and microscopy of WATs with hematoxylin-eosin(HE)staining was performed to observe the morphological changes of adipocytes.The expression of lipid metabolism related gene cell death-inducing DFF45-like effector C(CIDEC)in eWAT were detected by qPCR and Western blot.Results Compared with the control group,Nutlin-3a was found to promote the body weight(P<0.001),but no effect on glucose tolerance and insulin sensitivity in DIO mice.Nutlin-3a treatment decreased the size of adipocytes and fat deposition in adipose tissue and downregulated the mRNA and protein levels of CIDEC in eWAT.Conclusions Nutlin-3a inhibits the formation of lipid droplets by downregulating expression of CIDEC in white adipose tissue.
2.Effect of IL-22 Activating Nrf2 Pathway on the Colonic Barrier in Mice with Liver Fibrosis
Xing LIU ; Xiaojuan XU ; Yanfang WEI ; Hongyou YAN ; Junyan HUO ; Ke LI ; Cuiping XU
Herald of Medicine 2024;43(11):1733-1739
Objective To study the effect of IL-22 on the colonic barrier and its relationship with Nrf2 pathway in liver fibrosis mice.Methods The mice were divided into four groups:the control group(CON group),the model group(MOD group),the interleukin-22 group(IL-22 group),and the IL-22+ML385 group(ML385,an inhibitor of Nrf2),with 10 mice in each group,and the modeling cycle was 8 weeks.Liquid feed containing alcohol and carbon tetrachloride olive oil were given intraperitoneally in all groups except the CON group;IL-22 was given on top of this in the IL-22 group;and ML385 was injected intraperitoneally in the IL-22+ML385 group one hour before IL-22 treatment.At the end of modeling,the livers were stained with HE and Masson staining to clarify whether fibrosis occurred in the mice;the feces were collected to detect the cocci to bacillus ratio and observe the growth of intestinal flora;the colons were stained with HE staining,immunofluorescence and immunohistochemistry,and analyzed for the expression of tight junction proteins ZO-1,Occludin,and the Nrf2 pathway proteins(Nrf2,HO-1,and NQO1).The expression of these proteins was analyzed by immunohistochemistry.Results Compared with the CON group,mice in the MOD group showed significant fibrosis in the liver tissue,inflammatory cell infiltration in the colon tissue,and decreased expression of tight junction proteins(P<0.05).No overgrowth of various pathogenic bacteria was seen in fecal media.And there was no significant difference in the bulb-to-bar ratio.Compared with the MOD group,both liver and colon histopathologic damage were reduced in the IL-22 group,and tight junction protein expression was elevated,in addition,the expression levels of Nrf2,NQO1,and HO-1 were also elevated(P<0.05),whereas there was no significant change in the IL-22+ML385 group.Conclusion IL-22 improved the colonic barrier function in liver fibrosis mice,and the mechanism was related to the activation of Nrf2 anti-oxidative stress pathway.
3.Campylobacter fetus subsp. fetus ST20 isolated and identified from the bacteremia patient
Bei WEI ; Fen GAO ; Yue LIU ; Yan ZHONG ; Yanfang LI ; Deju QIN ; Jincheng ZHOU ; Xuebin XU
Chinese Journal of Laboratory Medicine 2024;47(9):1102-1105
A 72-year-old female patient was admitted to the emergency department of Qintang District People′s Hospital of Guigang City in August 2023 due to chills and fever, abdominal distension and pain, diarrhea, cough and shortness of breath for 1 day. She had a history of chronic obstructive and pulmonary heart disease, stage Ⅲ hypertension, and ceftazidime allergy. Clinical diagnosis of acute bacterial infection of chronic obstructive pneumonia was made and levofloxacin combined with piperacillin/tazobactam were given as symptomatic treatment. The blood culture reported Campylobacter fetus after four days, and the patient was cured and discharged after seven days with negative blood culture. The morphology and mass spectrometry identification of the strain were consistent with the definition of Campylobacter fetus. Whole genome sequencing predicted the multi-site sequence type as Campylobacter fetus subsp. fetus( Cff) ST20, carrying the tetracycline resistance gene tet (O/M/O), 18 flagella genes (including rpoN gene from Campylobacter jejuni. these genes were not found in the other two Campylobacter fetus subspecies), and six virulence genes (including like-typhoidal toxin and typhoid toxin genes). The pathogen has the ecological characteristics of parasitic farmed animal colonization and the biological characteristics of high mobility and virulence. These attributes facilitated its entry into the bloodstream via the fecal-oral route, leading to invasive infections.
4.Compound Danshen Dripping Pill inhibits hypercholesterolemia/atherosclerosis-induced heart failure in ApoE and LDLR dual deficient mice via multiple mechanisms.
Yanfang YANG ; Ke FENG ; Liying YUAN ; Yuxin LIU ; Mengying ZHANG ; Kaimin GUO ; Zequn YIN ; Wenjia WANG ; Shuiping ZHOU ; He SUN ; Kaijing YAN ; Xijun YAN ; Xuerui WANG ; Yajun DUAN ; Yunhui HU ; Jihong HAN
Acta Pharmaceutica Sinica B 2023;13(3):1036-1052
Heart failure is the leading cause of death worldwide. Compound Danshen Dripping Pill (CDDP) or CDDP combined with simvastatin has been widely used to treat patients with myocardial infarction and other cardiovascular diseases in China. However, the effect of CDDP on hypercholesterolemia/atherosclerosis-induced heart failure is unknown. We constructed a new model of heart failure induced by hypercholesterolemia/atherosclerosis in apolipoprotein E (ApoE) and LDL receptor (LDLR) dual deficient (ApoE-/-LDLR-/-) mice and investigated the effect of CDDP or CDDP plus a low dose of simvastatin on the heart failure. CDDP or CDDP plus a low dose of simvastatin inhibited heart injury by multiple actions including anti-myocardial dysfunction and anti-fibrosis. Mechanistically, both Wnt and lysine-specific demethylase 4A (KDM4A) pathways were significantly activated in mice with heart injury. Conversely, CDDP or CDDP plus a low dose of simvastatin inhibited Wnt pathway by markedly up-regulating expression of Wnt inhibitors. While the anti-inflammation and anti-oxidative stress by CDDP were achieved by inhibiting KDM4A expression and activity. In addition, CDDP attenuated simvastatin-induced myolysis in skeletal muscle. Taken together, our study suggests that CDDP or CDDP plus a low dose of simvastatin can be an effective therapy to reduce hypercholesterolemia/atherosclerosis-induced heart failure.
5.Spontaneous intracranial hypotension caused by high-flow spinal cerebrospinal fluid leaks: analysis of 6 patients
Yanfang DAI ; Zhen WANG ; Zheng WANG ; Tengda LIU ; Kang WANG ; Chong SHEN ; Yan LI ; Jie WU ; Dezhou QI ; Tianxinyu XIA ; Hong YE ; Junjie LI ; Liyong WU
Chinese Journal of Neurology 2023;56(2):178-186
Objective:To analyze the clinical features of 6 patients with spontaneous intracranial hypotension caused by high-flow spinal cerebrospinal fluid leaks.Methods:The clinical characteristics, auxiliary examinations, treatment, and outcomes in 6 patients of spontaneous intracranial hypotension caused by high-flow spinal cerebrospinal fluid leaks enrolled in the Xuanwu Hospital, Capital Medical University from February 2021 to April 2022 were retrospectively reviewed.Results:All the 6 patients had orthostatic headaches. Brain magnetic resonance imaging showed dural enhancement and brain sagging and magnetic resonance myelography showed longitudinal extradural collection in all the patients. The high-flow spinal cerebrospinal fluid leaks were demonstrated in upper thoracic segments by the dynamic myelography. The headache disappeared after conservative treatment in 2 patients and treatment with targeted epidural blood patch in 4 patients.Conclusions:The diagnosis of spontaneous intracranial hypotension caused by high-flow spinal cerebrospinal fluid leaks with typical orthostatic headache and brain magnetic resonance imaging and myelography findings is not difficult. However, the localization of the site of high-flow spinal cerebrospinal fluid leaks in spontaneous intracranial hypotension depends on the dynamic myelography. Targeted epidural blood patch is effective, but conservative treatment does not always work.
6.Spontaneous intracranial hypotension: analysis of 118 patients ' clinical characteristics
Zhen WANG ; Tianxinyu XIA ; Hong YE ; Jie WU ; Dezhou QI ; Zheng WANG ; Tengda LIU ; Chong SHEN ; Yan LI ; Yanfang DAI ; Zhongyun CHEN ; Junjie LI ; Liyong WU
Chinese Journal of Neurology 2023;56(9):1001-1008
Objective:To summarize the clinical features, radiological characteristics, therapy, and outcome of patients with spontaneous intracranial hypotension (SIH).Methods:The general information, clinical manifestations, auxiliary examinations, treatment, and outcomes in consecutive patients of SIH hospitalized in the Xuanwu Hospital, Capital Medical University from November 2018 to October 2022 were analyzed.Results:A total of 118 patients with a female-to-male ratio of 5∶4 were included and the ages were 17.00-71.00[39.00(34.00,46.75)]years with a preponderance in the age of 30-49 years. Almost all patients had orthostatic headaches (117/118, 99.2%), accompanied by nausea (90/118, 76.3%), vomiting (70/118, 59.3%), neck stiffness (88/118, 74.6%), tinnitus (57/118, 48.3%), and ear fullness (57/118, 48.3%). Brain magnetic resonance imaging (MRI) showed dural enhancement (97/113, 85.8%), enlarged venous sinus (88/113, 77.9%), subdural fluid collection (46/113, 40.7%), decreased suprasellar cistern (86/113, 76.1%), effacement of the prepontine cistern (86/113, 76.1%), diminished mamillopontine distance (80/113, 70.8%). The cerebrospinal fluid (CSF) leaks were detected in 90.7% (107/118) of the patients by magnetic resonance myelography but 54.3% (25/46) and 52.6% (20/38) by CT myelography and magnetic resonance myelography with gadolinium. Lumber puncture found CSF pressure<60 mmH 2O (1 mmH 2O=0.009 8 kPa) in 18.4% (19/103) of patients, increased CSF red blood cell counts in 50.6% (44/87) of patients, CSF pleocytosis in 44.8% (39/87) of patients, increased CSF protein concentrations in 57.5% (50/87) of patients. The headache completely disappeared after conservative treatment in 24.6% (31/118) of patients and after a single targeted epidural blood patch in 89.7% (78/87) of patients. A rebound headache after epidural blood patch treatment occurred in 66.0% (58/87) of patients. Conclusions:The patients with SIH almost manifested with orthostatic headache, and brain MRI and magnetic resonance myelography were suggested in those patients instead of CSF pressure by lumber puncture. Targeted epidural blood patch was effective and safe in SIH patients.
7.Rhein Attenuates Migration of Breast Cancer MDA-MB-231 Cells Through Inhibiting Mitochondrial Fission and EMT
Xiaobo LI ; Yan XIE ; Jinxia CHEN ; Xingfeng LOU ; Chunlong MIAO ; Qing FAN ; Yanfang ZHENG
World Science and Technology-Modernization of Traditional Chinese Medicine 2023;25(8):2692-2698
Objective To investigate the effects of the rhein on the mitochondria fission and epithelial-mesenchymal transition(EMT)of breast cancer cells MDA-MB-231.Methods Human breast cancer cells were intervened with rhein,and the cells were divided into control group(0 μmol·L-1),low dose rhein group(100 μmol·L-1),and high dose rhein group(200 μmol·L-1).The proliferation activity of the cells was detected by CCK-8,and migrations was detected by Scratch-healing migration assay.The morphology and distribution of mitochondria were detected by transmission electron microscope,and the expression levels of Dynamin-related protein 1(Drp1),mitofusin2(Mfn2),E-cadherin,Vimentin proteins were detected by Western blot.Results Compared with control group,Rhein significantly reduced the protein expression of Drp1、Vimentin(P<0.05),and increased E-cadherin and Mfn2,thus down-regulating mitochondria fission,inhibiting cell proliferation and migration.High dose Rhein was better than low dose.Conclusion Rhein can inhibit the proliferation and migration of breast cancer cells by reducing the expression of Drp1,Vimentin and up-regulating Mfn2,E-cadherin proteins.
8.Investigation on the Influence of Physician Experience on the Diagnosis and Treatment of Polycystic Ovary Syndrome
Yue WANG ; Jie CHEN ; Yan DENG ; Yanfang WANG ; Xiao MA ; Xuesong DING ; Jingwen GAN ; Yingying GUO ; Aijun SUN
Journal of Practical Obstetrics and Gynecology 2023;39(12):917-922
Objective:To investigate the differences in understanding and diagnosis of polycystic ovary syn-drome(PCOS)among obstetricians and gynecologists with different clinical experiences,in order to promote com-munication and improve medical quality.Methods:This was a multi-center cross-sectional survey.Information on participants'basic information and their knowledge of PCOS comorbidities and treatment management was col-lected online.Physicians were divided into two groups according to their experience:experienced(>200 consulta-tions/year)and inexperienced(≤200 consultations/year).The differences in the diagnosis and treatment of PCOS among different groups were compared.Logistic regression analysis was used to explore other factors that might influence the diagnosis and treatment of PCOS.Results:1689 questionnaires were finally included,among which 95 were experienced and 1594 were inexperienced.①In the comparison of basic conditions,the proportion of experienced persons in tertiary hospitals and reproductive endocrinology specialties was higher than that of in-experienced persons(P<0.05);The percentage of experienced persons who predicted patients with comorbid insulin resistance(proportion>30%)and predicted comorbid overweight/obesity(proportion 30%-50%)was higher than that of less experienced persons(P<0.05).②There was no significant difference in the evaluation of PCOS and its complications among different groups(P>0.05).Logistic regression suggested that different expe-riencers did not significantly affect the assessment of androgens,OGTT and lipid testing(P>0.05).③There was no statistical significance in the choice of treatment for those with no fertility need(P>0.05).For those with fertili-ty needs,metformin and letrozole were more actively selected by experienced group than inexperience(P<0.05).In terms of weight loss,experienced participants were more active in using metformin(P<0.05)and were more likely to choose1.0 or1.5 g/d(P<0.05)than those with inexperience.Conclusions:There was no signifi-cant difference in PCOS assessment among participants with different experience,but in treatment,experienced participants were better able to take into account the multi-system abnormalities brought by PCOS,and at the same time,they give more comprehensive and accurate treatment opinions on patients with or without fertility needs.
9.Effects of different hemostasis methods in laparoscopic ovarian cyst surgery on postoperative pain and sex hormone levels
Yan GUO ; Jiezhi ZHU ; Shuangling JIN ; Yanfang GAO
Chinese Journal of Endocrine Surgery 2023;17(3):327-331
Objective:To study the effect of different hemostasis methods on postoperative pain and sex hormone levels in patients undergoing laparoscopic ovarian cystectomy.Methods:A total of 118 patients with ovarian cysts admitted to our hospital from Jun. 2018.6 to Aug. 2020 were collected and grouped by digital table method into electrocoagulation hemostasis group (59 cases, electrocoagulation hemostasis) and suture hemostasis group (59 cases, suture hemostasis). Pain at time points, serum inflammatory factors and sex hormone levels in the two groups were measured, and the incidence of complications was counted 12 weeks after surgery.Results:The VAS scores of suture hemostasis group at 6, 12 and 24 h after operation (3.33±0.93, 3.63±1.02, 3.01±0.94) were significantly lower than those of the electrocoagulation hemostasis group (4.16±1.05, 4.61±1.17, 3.72±1.05) ; there was no significant difference in serum interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) levels between the preoperative suture hemostasis group and the electrocoagulation hemostasis group. The serum levels of IL-6 and TNF-α in the group (37.64±4.82ng/L, 39.67±4.71ng/L) were lower than those in the electrocoagulation hemostasis group (45.96±5.25ng/L, 48.96±5.14ng/L) ( P<0.05) .) ; there was no significant difference in serum follicle-stimulating hormone (FSH), estradiol (E 2) or luteinizing hormone (LH) levels between preoperative electrocoagulation hemostasis group and suture hemostasis group ( P>0.05) ; There was no significant difference in the three indexes in the suture hemostasis group 3 months after operation compared with those before treatment ( P>0.05). The serum FSH and LH [ (6.59±0.91) mIU/ml, (5.24±0.77) mIU/ml] in the suture hemostasis group were lower than those in the electrocoagulation hemostasis group [ (7.39±1.02) mIU/m, (5.97±0.89) mIU/m], E 2 in suture hemostasis group [ (51.08±6.09) pg/ml] was higher than that in electrocoagulation hemostasis group [ (46.88±5.59) pg/ml] ( P<0.05). In terms of the complication rate at 3 months after operation, the suture hemostasis group (32.20%) was significantly lower than electrocoagulation hemostasis (13.56%) ( P<0.05). After 1 year of follow-up, the pregnancy success rate of the suture hemostasis group (72.88%) was significantly higher than that of the electrocoagulation hemostasis group (52.54%) ( P<0.05). There was no significant difference in pregnancy outcomes ( P>0.05) . Conclusions:Suture hemostasis in patients undergoing laparoscopic ovarian cystectomy is beneficial to relieve postoperative pain, improve postoperative inflammatory response, protect their ovarian function, avoid complications such as abnormal ovulation and excessive menstrual flow, and improve the success rate of pregnancy. The overall application effect is better than electrocoagulation hemostasis.
10.Expression and significance of response gene to complement 32 in liver regeneration after partial hepatectomy in mice
Xingyuan LI ; Yanfang YANG ; Yan CHEN ; Wenhui HU ; Xiaoying ZHAO ; Junming TANG ; Deying KONG
Journal of Clinical Hepatology 2023;39(10):2396-2405
ObjectiveTo investigate the expression and role of response gene to complement 32 (RGC32) in liver regeneration after partial hepatectomy (PH). MethodsA total of 42 male C57BL/6 mice, aged 10 weeks, were randomly divided into control group, postoperative day 1 group (1-d group), postoperative day 2 group (2-d group), postoperative day 4 group (4-d group), postoperative day 6 group (6-d group), postoperative day 8 group (8-d group), and postoperative day 10 group (10-d group), with 6 mice in each group. In the control group, the complete liver of the mice was resected for weighing and photography as the normal control group (sham group); further, the left and middle lobes of the liver were resected for weighing and photography as the surgical control group (0-day group); the sham group and the 0-day group shared the same group of mice. After successful modeling by PH, the mice were sacrificed on days 1, 2, 4, 6, 8, and 10 after surgery, and the liver was collected to measure the change in size. HE staining and oil red O staining were used to evaluate liver histomorphological changes; serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured to evaluate the changes in liver function; immunohistochemical staining was used to measure the expression of proliferating cell nuclear antigen (PCNA) and Ki67 and analyze the change in cell proliferation during liver regeneration; quantitatie real-time PCR and immunohistochemical staining were uused to measure the expression and subcellular distribution of RGC32 during liver regeneration; EdU cell proliferation assay was used to analyze the effect of RGC32 overexpression or knocknout on hepatocyte proliferation in L02 cells. For continuous data, comparison between multiple groups was made by analysis of variance, and further pairwise comparisons were conducted using the LSD-t test. The independent samples t-test was used for comparison of continuous data between two groups. A Pearson correlation analysis was performed. ResultsThe liver gradually enlarged after PH, and the liver/body weight ratio rose to the peak from days 0 to 6, with significant differences between different time points (all P<0.05), while there was no significant change in liver size from days 6 to 10. The number of liver lipid droplets significantly increased after PH surgery and gradually decreased with liver regeneration, with a significant difference between the portal vein region and the central vein region (all P<0.05). Compared with the sham group, the 1d group had significant increases in the serum levels of ALT and AST (all P<0.05), which gradually returned to the levels of the sham group on day 6 and day 2 after surgery, respectively (P>0.05). Immunohistochemical staining showed that there were rapid increases in the numbers of PCNA- and Ki67-positive liver parenchymal cells after PH surgery, with the highest numbers of 86±5 and 89±5, respectively, on day 2, which then gradually decreased; however, there were gradual increases in the numbers of PCNA- and Ki67-positive nonparenchymal cells, with the peak numbers of 34±5 and 25±3, respectively, on day 6, which then gradually decreased. The total expression of RGC32 increased to the highest level on day 2 after PH surgery and then gradually decreased, and the changing trend of RGC32 expression in cytoplasm was consistent with that of total RGC32 expression; however, the expression of RGC32 in nucleus decreased to the lowest level on day 2 after PH surgery and then increased gradually. The correlation analysis showed that the expression of RGC32 in nucleus was negatively correlated with the proliferation of liver parenchymal cells (R2=0.308 3, P=0.016 7), and the expression of RGC32 in cytoplasm was positively correlated with the proliferation of liver parenchymal cells (R2=0.808 6, P<0.000 1). Cell experiments showed that compared with the control group, the EdU-positive rate was reduced by 15.6% after RGC32 overexpression (P<0.01) and was increased by 19.2% after RGC32 knockdown (P<0.01). ConclusionLiver parenchymal cells and nonparenchymal cells show asynchronous proliferation and participate in liver regeneration together. During liver regeneration after hepatectomy, there are differences in the expression of RGC32 between nucleus and cytoplasm, and RGC32 in nucleus may inhibit hepatocyte proliferation.

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