1.Effects of lncRNA MIR22HG on proliferation,apoptosis and inflammatory response of rheumatoid arthritis fibroblast -like synoviocytes by sponge adsorption of miR - 22-5p
Zhou Yang ; Shudian Lin ; Yuwei Zhan ; Lu Xiao ; keying Fu ; Xiaodie Huang
Acta Universitatis Medicinalis Anhui 2023;58(3):405-412
Objective:
To explore the effects of long non-coding RNA (LncRNA) MIR22HG on proliferation,apoptosis and inflammatory response of rheumatoid arthritis (RA) fibroblast-like synoviocytes (FLSs) and its molecular mechanism.
Methods:
Synovial tissue samples were collected from 37 RA patients and 30 joint trauma patients in our hospital,and the expression levels of MIR22HG and miR-22-5p in synovial tissue were detected by qRT-PCR. RA-FLSs in human was isolated ,cultured and identified in vitro. MIR22HG siRNA interference plasmid (si-MIR22HG) and its negative control plasmid (si-NC) ,miR-22-5p inhibitor and its negative control (inhibitorNC) were transfected into RA-FLSs respectively or simultaneously.The expression levels of MIR22HG and miR- 22-5p were detected by qRT-PCR. CCK-8 was used to detect the proliferation activity of cells in various groups. Annexin Ⅴ- FITC / PI was used to detect the apoptosis rates of cells in various groups.ELISA was used to detect the levels of TNF-α , IL-1 β and IL-6 in the supernatant of cells in various groups.Western blot was used to detect the protein expression levels of Bcl-2,Bax and Cleaved caspase-3 of cells in various groups.The targeting relationship between MIR22HG and miR-22-5p was verified by dual luciferase reporter gene assay.
Results :
Compared with joint trauma patients,the expression level of MIR22HG in synovial tissues of RA patients increased (P<0. 05) , while the expression level of miR-22-5p decreased (P<0. 05) .Interference with MIR22HG inhibited the proliferation activity of RA-FLSs,decreased the levels of TNF-α , IL-1 β and IL-6 in cell supernatant and the protein expression level of Bcl-2 in cells (P<0. 05) ,and increased the apoptosis rate,the expression level of miR-22-5p and the protein expression levels of Bax and Cleaved casepase-3 (P <0. 05 ) .However,inhibition of miR-22-5p expression reversed the effects of MIR22HG gene silencing on proliferation,apoptosis and inflammation of RA-FLSs (P<0. 05) .Dual luciferase reporting assay showed that miR-22-5p was a potential downstream miRNA target of MIR22HG.
Conclusion
MIR22HG is highly expressed in synovial tissues of RA patients,and it may promote the proliferation and the inflammatory response of RA-FLSs and inhibit cell apoptosis by down regulating the expression of miR-22-5p.
2. Establishment of population pharmacokinetics model of vancomycin in patients with Neutropenia
Liangmo LIN ; Jian XIAO ; Liangmo LIN ; Xiangjun FU ; Lili ZHONG ; Hefang WANG ; Qiongshi WU ; Jian XIAO
Chinese Journal of Clinical Pharmacology and Therapeutics 2021;26(9):1014-1022
AIM: To establish a population pharmacokinetics (PPK) model of vancomycin in patients with hematological diseases who developed neutropenia. METHODS: Patients from department of hematology with neutropenia in our hospital were taken into oue study.The patients (n=77) were performed trough and peak serum concentration of vancomycin, and their clinical data and medication information were collected. The Nonlinear mixed effect modeling approach (NONMEM) was used to establish the PPK model of those patients and model assessment and validation was carried out. Goodneess of fit plots and visual predictive check plus Bootstrap approach were used to assess validate our model. RESULTS: The model was a two compartment model, the final formulas were: clearance rate CL=6.84×(BW/70)
3.Effect of LncRNA GAPLINC on the Cell Proliferation of Rheumatoid Arthritis Fibroblast-like Synoviocytes
Bi-yao MO ; Lu XIAO ; Yu-wei ZHAN ; Yang YANG ; Li LIU ; Shu-dian LIN
Journal of Sun Yat-sen University(Medical Sciences) 2023;44(5):792-800
ObjectiveTo investigate the effect of LncRNA GAPLINC on the cell proliferation of RA-FLSs. MethodsRA-FLSs were cultured from synovial specimens. The expression of LncRNA GAPLINC in RA-FLSs and trauma-FLSs groups was detected by qRCR. GAPLINC suppression was transfected by siRNA and the inhibition efficiency was detected by qRCR. Flow cytometry was adopted to determine the change of cell growth and cell cycle distribution. 【ResμLts】 The expression of LncRNA GAPLINC was significantly higher in RA-FLSs than that of the trauma-FLSs (P<0.05).Transfection of GAPLINC-siRNA significantly decreased the expression of LncRNA GAPLINC. GAPLINC silence in RA-FLSs revealed significant inhibition in cell proliferation which was showed by the reduced cell number in S phase(P<0.05). Moreover, flow cytometry assay showed GAPIINC-siRNA treatment group had an accumμLation of cells in the G0/G1 phase and decreased RA-FLSs in the S and G2/M phase(P<0.05). After GAPLINC knockdown, mRNA and protein levels of Cyclin D1 and PCNA, which were positively correlated with proliferative phenotype, were decreased (P<0.05), while p21, which was negatively correlated with proliferative phenotype, was up-regμLated (P<0.05). ConclusionsThe mRNA expression of GAPLINC was higher in RA-FLSs compared with trauma-FLSs ,which was statistically significant(P<0.05). The silence of LncRNA GAPLINC coμLd significantly inhibit RA-FLSs cell growth and suppress the cell cycle transformation, which suggests that GAPLINC may play a role in the regμLation of proliferation of RA-FLSs, leading to synovial hyperplasia and contributing to RA progression.
4.Influence of bandage contact lens on corneal refractive status and postoperative complications in patients with pterygium
International Eye Science 2021;21(5):919-922
AIM: To analyze the influence of bandage contact lens on corneal refractive status and postoperative complications in patients with pterygium.
METHODS: Totally 116 patients with pterygium treated in the hospital between January 2018 and December 2019 were recruited in the study. They were divided into control group and observation group by random number table method, 58 cases in each group. The control group received extended pterygium resection and autologous corneal limbal stem cell transplantation. On this basis, the observation group was treated with bandage contact lens. The best corrected visual acuity(BCVA), surface asymmetry index(SAI)of corneal topography, surface regularity index(SRI), corneal refractive power and postoperative complications in the two groups were compared.
RESULTS: Compared with preoperation, the BCVA of the two groups was significantly higher 1mo and 3mo after operation(P<0.05), without statistically significant difference between the groups(P>0.05). Corneal horizontal and vertical curvature of the two groups were significantly higher, while corneal astigmatism was significantly lower at 1mo and 3mo after surgery. Meanwhile, the observation group had significantly lower corneal astigmatism than the control group(P<0.05). SAI and SRI of the two groups significantly decreased, which were significantly lower in the observation group than in the control group(P<0.05). No complications such as infection, poor healing of grafts and subconjunctival cysts were observed. The incidence of eye irritation was significantly lower in the observation group than in the control group, and lower at 2wk than at 1wk before operation(P<0.05).
CONCLUSION: Extended pterygium resection combined with autologous corneal limbal stem cell transplantation and bandage contact lens is more conductive to the improvement of corneal refraction in patients with pterygium, which also can reduce the incidence of eye irritation.
5.Analysis on identification and traceability of one non-toxigenic Corynebacterium diphtheriae from a patient with diabetic foot in Hainan province.
Xiao Jun ZHOU ; Cun Ren CHEN ; Xu Ming WANG ; Hua WU ; Tao HUANG ; Shao Ling WANG ; Lina NIU
Chinese Journal of Preventive Medicine 2022;56(8):1107-1111
There is a rare case of an elderly diabetic with diabetic foot infection at Hainan General Hospital in September 2021, which was diagnosed as Corynebacterium diphtheriae infection incidentally on routine culture with conventional methods and molecular biological approaches, to aid in diagnosis in clinical practice. Owing to smear staining, Albert staining and VITEK 2 system, automated identification systems viz matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) confirmed combing with 16S ribosomal RNA (16S rRNA) gene has been used for the taxonomic classification of bacteria. Otherwise, toxin gene tox was done for diphtheria toxin synthesis. The isolate was Gram-stain-positive, rod-like arrangement with irregular thickness, with characteristic metachromatic granules, ferment most sugars and homology of 16S rRNA analyses with C. diphtheriae NCTC11397T (MW682323.1) was greater than a 100% possibility, toxin gene tox was negative. The findings lay the foundation to clinical identify and trace of non-toxigenic C. diphtheriae. Moreover, this work provides insights into the non-toxigenic C.diphtheriae that contribute to recognized risk of non-toxigenic C.diphtheriae infections.
Aged
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Corynebacterium/genetics*
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Corynebacterium diphtheriae/genetics*
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Diabetes Mellitus
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Diabetic Foot
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Diphtheria/microbiology*
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Humans
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RNA, Ribosomal, 16S/genetics*
6. Clinical analysis of nine cases with critical corona virus disease 2019 in Hainan province
Ming LIU ; Feng LIN ; Jiao WANG ; Chaochao WEI ; Jia TIAN ; Juan FU ; Shaohua ZHONG ; Xinping CHEN ; Lizhen HAN ; Hui LI ; Jing CAO ; Suoxian CHEN ; Furong XIAO ; Yongxing CHEN ; Zhongyi ZHOU ; Xiaohong XIE ; Tao WU
Chinese Journal of Infectious Diseases 2020;49(0):E024-E024
Objective To explore the clinical features of critical cases of coronavirus disease 2019 (COVID-19). Methods The clinical data of nine patients who were diagnosed with critical COVID-19 in Hainan General Hospital from January 21, 2020 to February 6, 2020 were retrospectively analyzed. RT-PCR testing for 2019 novel coronavirus (2019-nCoV) was performed with multi-sites synchronize specimens including pharyngeal swab, blood, excrement, and urine. The serum levels of leucocyte, C-reactive protein, procalcitonin and lactic acid between the improved group (five cases) and the deteriorated group (four cases) were compared. The t test was used for comparison of normally distributed continuous data between groups. Results There were eight males (88.9%) and 1 female enrolled. The patients aged 28-77 years old, with an age of (52.9±18.0) years. By March 4, 2020, all five cases in improved group were cured and discharged, three cases in deteriorated group died and 1case remained in critical condition. All multi-sites specimens of patients in improved group turned negative in 2-4 weeks of illness onset, while those of cases in deteriorated group showed sustained viral nucleic acid positive (up to 48th day of illness onset). The white blood cell counts ((13.52±8.24)×10 9 /L vs (10.49±4.46) ×10 9 /L), C-reactive protein ((139.71±87.46) mg/L vs (78.60±55.40) mg/L) and procalcitonin ((2.32±4.03) ng/mL vs (0.28±0.58) ng/mL) , lactic acid ((3.70±4.14) mmol/L vs (2.33±0.53) mmol/L) in deteriorated group were all significantly higher than those in improved group ( t =2.908, 5.009, 4.391 and 2.942, respectively, all P <0.01). A rapid rise of serum IL-6 level up to 8 500 pg/mL was observed in one patient three days prior to death. Conclusion Among the patients with critical COVID-19, serum levels of inflammatory cytokines of the death cases are higher than those of improved and discharged cases.
7. Effect of retrograde colonic electrical stimulation on colonic transit and stress-induced visceral hypersensitivity in rats with irritable bowel syndrome
Xiao-Ri QIN ; Xiao-Ning SUN ; Yan TAN
Asian Pacific Journal of Tropical Medicine 2017;10(8):827-832
Objective To evaluate the effects of retrograde colonic electrical stimulation (RCES) with trains of short pulses and RCES with long pulses on colonic transit in irritable bowel syndrome (IBS) rats and to investigate whether stress-induced visceral hypersensitivity could be alleviated by RCES so as to find a valuable new approach for IBS treatment. Methods A total of 48 male rats were randomly divided into model group and control group. Visceral hypersensitivity model was induced by a 6-day HIS protocol composed of two stressors, restraint stress for 40 min and forced swimming stress for 20 min. The extent of visceral hypersensitivity was quantified by electromyography and abdominal withdrawal reflex scores (AWRs) of colorectal distension (use a balloon) at different pressures. After the modeling, all rats were equipped with electrodes in descending colon for retrograde electrical stimulation and a PE tube for perfusing phenol red saline solution in the ileocecus. After recovering from surgery, RCES with long pulses, RCES with trains of short pulses, and sham RCES were performed in colonic serosa of rats for 40 min in six groups of 8 each, including three groups of visceral hypersensitivity rats and three groups of health rats. Colonic transit was assessed by calculating the output of phenol red from the anus every 10 min for 90 min. Finally, the extent of visceral hypersensitivity will be quantified again in model group. Results After the 6-day HIS protocol, the HIS rats displayed an increased sensitivity to colorectal distention, compared to control group at different distention pressures (P < 0.01). CRES with trains of short pulses and long pulses significantly attenuated the hypersensitive responses to colorectal distention in the HIS rats compared with sham RCES group (P < 0.01). The effects of RCES on rats colon transmission: In the IBS rats, the colonic emptying were (77.4 ± 3.4)%, (74.8 ± 2.4)% and (64.2 ± 1.6)% in the sham RCES group, long pulses group and trains of short pulses group at 90 min; In healthy rats, The colonic emptying was (65.2 ± 3.5)%, (63.5 ± 4.0)% and (54.0 ± 2.5)% in the sham RCES group, long pulses group and trains of short pulses group at 90 min. Conclusion RCES with long pulses and RCES with trains of short pulses can significantly alleviate stress-induced visceral hypersensitivity. RCES with trains of short pulses has an inhibitory effect of colonic transit, both in visceral hypersensitivity rats and healthy rats.
8.Delayed K562 cell apoptosis promoted by cleaved LyGDI after 60Co γ-rays irradiation
Huali SUN ; Weiming DUAN ; Yanyan SHAO ; Hainan XIAO ; Xinwen ZHOU
Chinese Journal of Radiological Medicine and Protection 2010;30(6):643-646
Objective To elucidate the function and regulatory mechanism of LyGDI involved delayed cell death in the human K562 cells and HL-60 cells induced by 60Co γ-rays. Methods Erythrosine B cells staining was used to count the apoptosis rate. PI staining and flow cytometry were applied to check the cell cycle. The expression of LYGDI and Rac1 was resolved by Western blot by using monoclonal antibody of LyGDI and Racl. The distribution of Racl protein in cells was observed with immunofluorescence by using the confocal microscope. Results The K562 cells showed G2/M phase arrest and the percent age was 71.3%. The apoptosis rate was very low at early post-irradiation stage in the K562 cells. The apoptosis rate was 14% in the K562 cells at 24 h post-irradiation with 8 Gy of γ-rays, and delayed cell apoptosis was present. LyGDI was cleaved in the K562 cells irradiated by 4 Gy 60 Co γ-rays after 24 hours post-irradiation. The expression of Racl protein was not altered at all, but the distribution was changed in the irradiated cells while the Racl protein moved to cell membrane and a little in cell nucleus. The Racl was activated with the losing the binding affinity with the LyGDI. Conclusion LyGDI could promote the delayed cell apoptosis, which is through the activation of the Rac1.
9. Chemical constituents from root bark of Litsea glutinosa
Chinese Traditional and Herbal Drugs 2019;50(12):2817-2821
Objective: To study the chemical constituents from the root bark of Litsea glutinosa. Methods: Silica gel and Sephedex LH-20 column chromatographies as well as semi-preparative HPLC were applied to isolate and purify the compounds. Their structures were elucidated based on the spectrum analysis. Results: Fifteen compounds were obtained and identi-fied as schizandriside (1), lyonside (2), coclaurine (3), alangisesquin A (4), alangisesquin B (5), dihydrobuddlenol (6), ssioriside (7), dendranthemoside B (8), epi-anhydrocinnzeylanol (9), isolariciresinol-5’-methoxy-iso-larixa-9’-O-β-D-xylopyranoside (10), benzyl alcohol-β-D- glucopyranoside (11), phenylethyl-β-D-glucopyranoside (12), n-butyl-β-D-fructoside (13), N-cis-feruloyl tyramine (14), and N-trans-sphingoyl tyramine (15). Conclusion: Compounds 1-13 are isolated from this plant for the first time.