1.Effect of knockdown of ARHGAP30 on proliferation and apoptosis of Siha cells
Ya-Ting PENG ; Duan LIU ; Jie MENG ; Wen-Chao LI ; Hui-Qi LI ; Hua GUO ; Mei-Lan NIU ; Qiao-Hong QIN
Chinese Pharmacological Bulletin 2024;40(5):847-853
Aim To investigate the changes in the proliferation and apoptosis of Siha cells after knocking down Rho GTPase-activating protein 30(ARHGAP30).Methods After designing specific shARHGAP30 primers and connecting them to the pLKO.1 vector,we transformed them into Escherichia coli competent cells,then co-transfecting them with lentiviral helper plasmids into HEK-293T cells.We collected and filtered cell supernatant to obtain the vi-rus to infect Siha cells.RT-qPCR and Western blot were used to detect knockdown efficiency,as well as changes in the expression of Bax and Bcl-2 after trans-fection.The CCK-8 method was employed to measure the proliferation level of cells after knockdown.Results After successful construction of a lentiviral plasmid with knockdown of the ARHGAP30 gene and establish-ment of stably transfected Siha cells,ARHGAP30 tran-scription and translation(P<0.01)in Siha cells de-creased,Bax/Bcl-2 significantly decreased(P<0.01),indicating decreased apoptosis and increased cell proliferation(P<0.01).Conclusions This study suggests the involvement of ARHGAP30 in the proliferation and apoptosis of Siha cells,and regulating the ARHGAP30 gene may interfere with the occurrence and development of cervical cancer.
2.Heshouwuyin delaying the aging of Leydig cells in rat testis through DNA methyltransferase 1
Tian WU ; Shuo-Ya LU ; Yu-Jiao YANG ; Yu-Lei DUAN ; Qi-Chao AN ; Xiao-Lan ZHEN ; Si-Yun NIU
Acta Anatomica Sinica 2024;55(3):276-284
Objective To investigate whether Heshouwuyin can delay the aging of Leydig cells in rat testis through DNA methyltransferase 1(DNMT1).Methods Totally 40 male Wistar rats were randomly divided into 4 groups,with 10 rats in each group.Immunohistochemistry was used to detect the expression levels of DNMT1 in testis tissue of rats.Testosterone content in serum of rats in each group was detected by ELISA test.A rat Leydig cell aging model was established by free radical oxidative damage.DNMT1 was knocked down by lentivirus in Leydig cells,and the cell senescence status and the testosterone content and testosterone synthesis key enzyme 3β-hydroxysteroid dehydrogenase(3β-HSD),cytochrome P450 family member 11A1(CYP11A1)content secreted by cells were detected by β-galactosidase(β-GAL)staining,immunofluorescenct staining and ELISA.Results Compared with the young control group(YCG),the expression of P16 protein and the positive rate of β-GAL in the testis tissue of rats in the natural aging group(NAG)increased significantly,and the expression of DNMT1 and serum testosterone levels decreased(P<0.05).However,after Heshouwuyin intervention,the expression of P16 protein and the positive rate of β-GAL in the testis of aging rats were reduced,and DNMT1 expression and the serum testosterone levels increased(P<0.05).The same trend was observed in Leydig cells.Knockdown of DNMT1 in Leydig cells,β-GAL positivity and P16 protein expression increased significantly,and testosterone secretion and testosterone synthesis key enzymes 3β-HSD,CYP11A1 content from Leydig cells decreased significantly,compared with the normal control group(NCG)(P<0.05).When Heshouwuyin was added,the above phenomenon was improved.Conclusion Heshouwuyin can delay the aging of rat Leydig cells through DNMT1.
3.Epidemiological investigation on the local epidemic situation in Zhengzhou High-Tech Zone caused by SARS-CoV-2 Delta variant.
Yue Fei JIN ; Yue LI ; Jun Wei LI ; Zhuo Ya YAN ; Shuai Yin CHEN ; Xiao Min LOU ; Ke FAN ; Fan WU ; Yuuan Yuan CAO ; Fang Yuan HU ; Long CHEN ; Ya Qi XIE ; Cheng CHENG ; Hai Yan YANG ; Guang Cai DUAN
Chinese Journal of Preventive Medicine 2023;57(1):43-47
This study collected epidemic data of COVID-19 in Zhengzhou from January 1 to January 20 in 2022. The epidemiological characteristics of the local epidemic in Zhengzhou High-tech Zone caused by the SARS-CoV-2 Delta variant were analyzed through epidemiological survey and big data analysis, which could provide a scientific basis for the prevention and control of the Delta variant. In detail, a total of 276 close contacts and 599 secondary close contacts were found in this study. The attack rate of close contacts and secondary close contacts was 5.43% (15/276) and 0.17% (1/599), respectively. There were 10 confirmed cases associated with the chain of transmission. Among them, the attack rates in close contacts of the first, second, third, fourth and fifth generation cases were 20.00% (5/25), 17.86% (5/28), 0.72% (1/139) and 14.81% (4/27), 0 (0/57), respectively. The attack rates in close contacts after sharing rooms/beds, having meals, having neighbor contacts, sharing vehicles with the patients, having same space contacts, and having work contacts were 26.67%, 9.10%, 8.33%, 4.55%, 1.43%, and 0 respectively. Collectively, the local epidemic situation in Zhengzhou High-tech Zone has an obvious family cluster. Prevention and control work should focus on decreasing family clusters of cases and community transmission.
Humans
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SARS-CoV-2
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COVID-19
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Epidemics
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Incidence
4.Xixin Decoction improves learning and memory ability of SAMP8 by enhancing neuroprotective effect and inhibiting neuroinflammation.
En-Long ZHAO ; Yong-Chang DIWU ; Hu ZHANG ; Li-Qi DUAN ; Xin-Yue HAN ; Ya-Li WANG ; Yuan ZHOU
China Journal of Chinese Materia Medica 2023;48(18):5032-5040
This study aimed to explore the possible effect of Xixin Decoction(XXD) on the learning and memory ability of Alzheimer's disease(AD) model senescence-accelerated mouse-prone 8(SAMP8) and the related mechanism in enhancing neuroprotective effect and reducing neuroinflammation. Forty SAMP8 were randomly divided into a model group(10 mL·kg~(-1)·d~(-1)), a probiotics group(0.39 g·kg~(-1)·d~(-1)), a high-dose group of XXD granules(H-XXD, 5.07 g·kg~(-1)·d~(-1)), a medium-dose group of XXD granules(M-XXD, 2.535 g·kg~(-1)·d~(-1)), and a low-dose group of XXD granules(L-XXD, 1.267 5 g·kg~(-1)·d~(-1)). Eight senescence-accelerated mouse-resistant 1(SAMR1) of the same age and strain were assigned to the control group(10 mL·kg~(-1)·d~(-1)). After ten weeks of intragastric administration, the Morris water maze was used to test the changes in spatial learning and memory ability of mice after treatment. Meanwhile, immunofluorescence staining was used to detect the positive expression of receptor for advanced glycation end products(AGER), Toll-like receptor 1(TLR1), and Toll-like receptor 2(TLR2) in the hippocampal CA1 region of mice. Western blot was employed to test the protein expression levels of silencing information regulator 2 related enzyme 1(SIRT1), AGER, TLR1, and TLR2 in the hippocampus of mice. Enzyme linked immunosorbent assay(ELISA) was applied to assess the levels of Aβ_(1-42) in the hippocampus of mice and the levels of nuclear factor κB p65(NF-κB p65), NOD-like receptor protein 3(NLRP3), tumor necrosis factor-α(TNF-α), and interleukin-1β(IL-1β) in the serum and hippocampus of mice. Compared with the model group, XXD significantly improved the spatial learning and memory ability of SAMP8, increased the expression of neuroprotective factors in the hippocampus, decreased the levels of neuroinflammatory factors, and inhibited the expression of Aβ_(1-42). In particular, H-XXD significantly increased the expression of SIRT1 in the hippocampus of mice, reduced the expression levels of NF-κB p65, NLRP3, TNF-α, and IL-1β in the serum and hippocampus of mice, and decreased the expression of AGER, TLR1, and TLR2 in the hippocampus of mice(P<0.05 or P<0.01). XXD may improve the spatial learning and memory ability of AD model SAMP8 by enhancing the neuroprotective effect and inhibiting neuroinflammation.
Humans
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Neuroprotective Agents/therapeutic use*
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Sirtuin 1/metabolism*
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Toll-Like Receptor 2/metabolism*
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NF-kappa B/metabolism*
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Tumor Necrosis Factor-alpha/metabolism*
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Neuroinflammatory Diseases
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NLR Family, Pyrin Domain-Containing 3 Protein/metabolism*
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Toll-Like Receptor 1/metabolism*
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Alzheimer Disease/genetics*
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Hippocampus
5.Changes in the intestinal flora of rats under high altitude hypoxia
Gui-qin LIU ; Xue BAI ; Ya-bin DUAN ; Jun-bo ZHU ; Jian-xin YANG ; Qian WANG ; Yang ZHOU ; Wen-qi GU ; Xiang-yang LI
Acta Pharmaceutica Sinica 2021;56(4):1100-1108
The structure and diversity of the intestinal flora in rats exposed to high altitude hypoxia was investigated. Animal experiments strictly follow the regulations of Medical Laboratory Animal Ethics Committee of Qinghai University, School of Medicine. SD rats were randomly divided into a control group, a moderate altitude hypoxia group, and a high altitude hypoxia group. The pH value of the feces was measured and histopathological changes in the small intestine were determined by HE staining, and the intestinal flora were characterized by 16S rDNA high throughput sequencing technology on the 3rd, 7th, 15th, and 30th day of hypoxia exposure. Compared with the control group, the fecal pH value of rats in the moderate altitude hypoxia group and the high altitude hypoxia group was decreased significantly. The lamina propria and submucosa capillaries were slightly dilated and congested on the 3rd day in the moderate altitude hypoxia group. In the high altitude hypoxia group the submembrane capillaries were dilated and congested, the lamina propria of the mucosa showed mild edema, and the lymphatic vessels were dilated on the 7th day. The composition and diversity of intestinal flora in these rats changed significantly with prolonged exposure to the high altitude hypoxic environment. A total of 35 phyla, 87 classes, 205 orders, 337 families, 638 genera, and 256 species were annotated in the three groups of rats, including Firmicutes, Clostridia, Clostridiales, Ruminococcaceae, Akkermansia, and Lactobacillus_murinus. Compared with the control group, the intestinal flora of the hypoxic groups showed the most significant changes by the 15th day. There were 9 microbiota of gut microorganisms with relative abundance in the moderate altitude hypoxia group, of which Rikenellaceae_RC9_gut_group bacteria was the most common, there were 19 different microbiota of gut microorganisms with higher relative abundance in the high altitude hypoxia group, of which Ruminococcaceae bacteria was the most common. The results of this study indicate significant changes in the intestinal flora with high altitude hypoxia, and establish a foundation for further research on the initiation and development of diseases and drug metabolism in high altitude hypoxia.
6.Comparison of clinical effects of different anterior surgical methods for the treatment of single segment cervical disc herniation.
Ya-Yi FAN ; Liang DUAN ; Hong-Hai XU ; Jun LIU ; Jie QI ; Li-Qun GONG
China Journal of Orthopaedics and Traumatology 2021;34(4):354-359
OBJECTIVE:
To compare the clinical effects of different anterior surgical methods in treating single segment cervical disc herniation.
METHODS:
The clinical data of 46 patients with single-segment cervical disc herniation underwent surgical treatment from September 2013 to September 2018 were retrospectively analyzed. The patients were divided into three groups according to different surgical methods. Among them, 23 patients in the anterior percutanousendomic cervical dissection (APECD) group, there were 8 males and 15 females, aged (47±3) years old, prominent segments were C
RESULTS:
All 46 patients were followed up for 12 to 24 (17.57±3.15)months. The follow-up time of APECD, CDR, ACDF groups were (17.30±3.25), (17.80±3.16), (17.85±2.88) months, and operation time were (95.48 ±13.85), (58.50±7.09), (76.00±15.72) min, respectively, there were no significant differences in follow-up time and operation time between two groups(
CONCLUSION
The three anterior surgical approaches can achieve satisfactory clinical results for the treatment of single-segment cervical disc herniation. However, the improvement rate of the CDR group and the activity of the retained responsibility segment are better than those of the other two groups. APECD surgery may have recurrence.
Adult
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Cervical Vertebrae/surgery*
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Diskectomy
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Female
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Follow-Up Studies
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Humans
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Intervertebral Disc Degeneration/surgery*
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Male
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Middle Aged
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Retrospective Studies
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Spinal Fusion
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Total Disc Replacement
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Treatment Outcome
7.Spectrometric analyses of larotaxel and larotaxel liposomes quantification by high performance liquid chromatography.
Xue Qi LI ; Jian Wei LI ; Qiu Hong LI ; Yan YAN ; Jia Lun DUAN ; Yi Nuo CUI ; Zhan Bo SU ; Qian LUO ; Jia Rui XU ; Ya Fei DU ; Gui Ling WANG ; Ying XIE ; Wan Liang LU
Journal of Peking University(Health Sciences) 2019;51(3):467-476
OBJECTIVE:
Larotaxel is a new chemical structure drug, which has not been marketed worldwide. Accordingly, the standard identification and quantification methods for larotaxel remain unclear. The spectrometric analyses were performed for verifying weight molecular formula, molecular weight and chemical structure of larotaxel. Besides, a quantification method was developed for measuring larotaxel in the liposomes.
METHODS:
The molecular formula, molecular weight and chemical structure of larotaxel were studied by using mass spectrometry (MS), infra-red (IR), nuclear magnetic resonance (NMR) and ultraviolet-visible (UV-vis) spectrometric techniques. The absorption wavelength of larotaxel was investigated by UV-vis spectrophotometry full-wavelength scanning. Besides, a quantification method was developed by high performance liquid chromatography (HPLC), and then validated by measuring the encapsulation efficacy of larotaxel liposomes.
RESULTS:
The four spectral characteristics of larotaxel were revealed and the corresponding standard spectra were defined. It was confirmed that larotaxel had the structure of tricyclic diterpenoids, with the molecular formula of C45H53NO14, the molecular weight of 831.900 1, and the maximum absorption wavelength of 230 nm. The quantitative method of larotaxel was established by using HPLC with a reversed phase C18 column (5 μm, 250 mm×4.6 mm), a mobile phase of acetonitrile-water (75:25, volume/volume), and a detection wavelength of 230 nm. The validation study exhibited that the established HPLC method was stable, and had a high recovery and precision in the quantitative measurement of larotaxel in liposomes. In addition, a new kind of larotaxel liposomes was also successfully prepared. The particle size of the liposomes was about 105 nm, with an even size distribution. And the encapsulation efficiency of larotaxel in the liposomes was above 80%.
CONCLUSION
The present study offers reference standard spectra of larotaxel, including MS, IR, NMR, and UV-vis, and confirms the molecular formula, molecular weight and chemical structure of larotaxel. Besides, the study develops a rapid HPLC method for quality control of larotaxel liposomes.
Chromatography, High Pressure Liquid
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Liposomes
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Magnetic Resonance Spectroscopy
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Taxoids
8.Sharp injuries among health care workers in Xuzhou
Yi-Xue ZHANG ; Yu-Lian GU ; Qi JIANG ; Ya-Qi WANG ; Si-Chen DUAN ; Yi-Ping MAO
Chinese Journal of Infection Control 2018;17(11):1003-1007
Objective To investigate the occurrence and reporting of sharp injuries among health care workers (HCWs)at all levels of hospitals in Xuzhou City,provide evidences for formulating protective measures against sharp injuries and improving the reporting system.Methods From July to August 2016,13 hospitals in Xuzhou City were randomly selected by multi-stage stratified random sampling method,the general information,occurrence of sharp injuries,and reporting situation was performed questionnaire survey.Results A total of 2 694 valid question-naires were collected,incidence,case incidence,and reporting rate of sharp injuries were 10.32%,12.84%,and 30.64% respectively.Case incidence of sharp injuries among HCWs in primary,secondary,and tertiary hospitals were 44.83%,11.53%,and 12.52% respectively,case incidences of sharp injuries in different levels of hospitals were significantly different (χ2=55.148,P<0.001).The main opportunity for sharp injuries was when HCWs re-turned needle cap (79 cases,22.83%),the main device involving sharp injuries was hollow-bore needle (297 cases, 85.84%).Incidences of sharp injuries among HCWs receiving different training were significantly different (χ2=66.760,P<0.001).Conclusion Current situation of sharp injuries among HCWs in this region is not optimistic, there are some problems such as poor training efficacy,low reporting rate and low use rate of safety devices,effec-tive measures should be taken to establish an effective monitoring and tracking system for sharp injuries,so as to re-duce the occurrence of sharp injuries.
9.Efficacy of Extended Myectomy on Reducing Mitral Regurgitation in Patients With Hypertrophic Obstructive Cardiomyopathy
Ya-Jie TANG ; Yun-Hu SONG ; Long DENG ; Fu-Jian DUAN ; Yun LIU ; Jun RAN ; Ge GAO ; Yuan LI ; Qi LI
Chinese Circulation Journal 2018;33(6):585-590
Objectives:To analyze the impact of extended myectomy on reducing mitral regurgitation in patients with hypertrophic obstructive cardiomyopathy (HOCM). Methods:We retrospectively analyzed 480 consecutive HOCM patients who underwent surgical treatment by the same surgeon in our institution from October 2002 to July 2017. The efficacy of extended myectomy for reducing mitral regurgitation and left ventricular outflow tract (LVOT) obstruction were evaluated by echocardiography after surgery. Results:Among the 480 patients, 22 (4.6%) received concomitant mitral repair or replacement because of their intrinsic mitral diseases. In the remaining 458 (95.4%) patients without concomitant mitral valve surgery, 1 (0.2%) died at the 5th day after surgery because of infective shock, and another 4 (0.9%) lost to follow-up, a total of 453 (98.9%) patients underwent echocardiographic follow-up (median follow-up time:6 months [3, 12]). During follow-up, left ventricular out flow tract gradient was significantly decreased from (89.1±30.6) to (12.8±11.6) mmHg (P<0.001); the number of patients with systolic anterior motion (SAM) of mitral leaflets decreased from 451(98.5%) to 42 (9.3%) (P<0.001); 297 (64.8%) patients presented with moderate or severe mitral regurgitation before surgery, which decreased to 14 (3.1%) at follow-up (P<0.001); the multivariate regression analysis showed that patients with residual SAM were significantly associated with a higher incidence of moderate to severe mitral regurgitation during follow-up (odds ratio 30.334, 95% confidence interval:5.619-163.739, P<0.001). Conclusions:Extended myectomy, combined with dividing the anomalous links between mitral apparatus and septum, and trimming papillary muscles, yields satisfactory outcomes of relieving LVOT obstruction and reducing mitral regurgitation in most of patients with HOCM. Concomitant mitral valve surgery is rarely required unless the patient have intrinsic mitral valve disease.
10.Comparison of minimal inhibitory concentrations of ciprofloxacin against Pseudomonas aeruginosa induced by Pseudomonas quinolone signal and ciprofloxacin
Yan-Ru WANG ; Zhi-Qi ZHANG ; Ya-Ting GAO ; Xiao-Xia LI ; Jian-Bang KANG ; Li-Xia QIU ; Jin-Ju DUAN
The Chinese Journal of Clinical Pharmacology 2018;34(9):1115-1118
Objective To investigate minimal inhibitory concentration (MIC) of ciprofloxacin against Pseudomonas aeruginosa induced by Pseudomonas quinolone signal (PQS) and ciprofloxacin in vitro.Methods Clinical isolates of Pseudomonas aeruginosa sensitive to ciprofloxacin were collected and then induced ciprofloxacin with three concentrations of 0.5 × MIC,2 × MIC and 4 × MIC,and PQS with three concentrations of 10,40,80 μmol· L-1,respectively for five days.The agar dilution method was used to measure MICs of all strains before and after inductions to ciprofloxacin.The MICs to ciprofloxacin before and after inductions of the same induction scheme were analysed by repeated measures analysis of variance and Paired t-test was used to compare the MICs of two induced schemes.Results Twelve clinical isolates of Pseudomonas aeruginosa sensitive to ciprofloxacin were obtained,among them one was used as the quality control strain included.There had interaction between induction time and induction concentrations of PQS or ciprofloxacin (P <0.001 or P <0.05).MICs of strains to ciprofloxacin of two induced schemes had statistically significant difference (P < 0.05).Conclusion Under different concentrations of PQS,the trend of MIC values of ciprofloxacin to Pseudomonas aeruginosa varied by induction time.Under different concentrations of ciprofloxacin,MIC values tended to increase with the prolongation of induction time.The effects of two induction schemes on MIC of ciprofloxacin were different.

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