1.Association between PNPLA3 polymorphisms (rs738409) and non-alcoholic fatty liver disease
Guowei LIANG ; Donghua SHAO ; Jie LIU ; Meilin HE
Chinese Journal of Laboratory Medicine 2016;(1):45-49
Objective To develop a novel method for genotyping rs 738409 ( C >G ) single nucleotide polymorphism ( SNP ) , and explore the association between the rs 738409 genotypes and non-alcoholic fatty liver disease ( NAFLD ) .Methods Method establishment and analysis of genetic susceptibility.The principle of amplification refractory mutation system ( ARMS) and combined a TaqMan fluorogenic probe as signal report were used , by monitoring the difference of cycle threshold (ΔCt=C allele-special primer Ct values minus G allele-special primer Ct values ) between the two PCR reactions in a real-time PCR, the method for rs738409 genotyping was established ( ARMS-TaqMan) .618 subjects ( men:401;women: 217 ) , in an annual health check-up program from January 2011 to December 2014 in Aerospace Center Hospital , were performed for rs738409 genotyping by the ARMS-TaqMan assay.Fatty liver was diagnosed based on abdominal ultrasonography .The chi-square test and multiple logistic analyses were used to analyze the relationship of rs 738409 genotypes and NAFLD.Results The ΔCt by ARMS-TaqMan for rs738409 genotyping were -13.1 ±1.4 of CC alleles ( 243 cases ) , 0.01 ±0.45 of CG alleles ( 282 cases), and 12.7 ±1.9 of GG alleles (93 cases), respectively.The GG alleles frequency of rs738409 were significantly higher in patients with NAFLD compared with subjects without NAFLD (21.5%vs 12.3%,χ2=8.677, P =0.003).In comparison to subjects with CC alleles, the OR (95% confidence interval) adjusted for age, gender, total cholesterol, triglyceride, high density lipoprotein-cholesterol, low density lipoprotein-cholesterol, fasting blood glucose and body mass index was 1.35 (0.91-2.00) in subjects with CG alleles and was 2.21 ( 1.32 -3.71 ) in subjects with GG alleles ( P =0.013 ) .Variant rs738409 genotypes were associated with significant increased trend in alanine aminotransferase ( ALT) level from CC alleles, CG alleles to GG alleles (F=8.980, P<0.001), and in aspartate aminotansferase (AST) between GG alleles and CC alleles (F=6.491, P<0.001).Conclusions The novel ARMS-TaqMan assay had the features of accuracy , one step and high-throughput for rs738409 genotyping.The G allele of rs738409 was a risk factor of NAFLD susceptibility and associated with higher level serum ALT and AST .
2.Promoting effect of AGTR1 blocker olmesartan on the apoptosis of HTF and its mechanism
Lijun WANG ; Hongsong LI ; Wenyi ZHANG ; Meilin SHAO ; Meimei REN ; Jianming WANG
Chinese Journal of Experimental Ophthalmology 2023;41(2):119-126
Objective:To study the effect and mechanism of angiotensin type 1 receptor (AGTR1) blocker olmesartan (OMS) on the apoptosis of human Tenon capsule fibroblasts (HTF).Methods:Tenon capsule tissues were obtained from patients during strabismus surgery in the Second Affiliated Hospital of Xi'an Jiaotong University.Primary HTF were cultured by explant culture.Primary cells were identified by vimentin immunofluorescence staining and flow cytometry.The fibrosis model of HTF was established using 10 ng/ml transforming growth factor-β2 (TGF-β2). The cells were divided into normal control group cultured in culture medium, TGF-β2 group in culture medium containing TGF-β2, TGF-β2+ OMS group in culture medium containing TGF-β2 and OMS, and OMS group in culture medium containing OMS, and were cultured for 48 hours.Cell apoptosis was detected by flow cytometry with annexin V/PI staining.The early apoptosis, late apoptosis, and total apoptosis rates were analyzed.The protein expression of procaspase-9, cleaved caspase-9, bax and bcl-2 in the mitochondrial apoptosis pathway was detected by Western blot.The activity of lactate dehydrogenase (LDH) and superoxide dismutase (SOD) was detected by colorimetry.The study protocol was approved by the Ethics Committee of the Second Affiliated Hospital of Xi'an Jiaotong University (No.2019-014).Results:Primary HTF were successfully isolated and cultured.The cultured cells were long spindle-shaped and positive for vimentin.The expression rate of vimentin in the primary cells was greater than 99%.A statistically statistical difference was found in the early apoptosis rate, late apoptosis rate, and total apoptosis rate among the four groups ( F=24.92, 3.96, 41.82; all at P<0.05). The early and total apoptosis rates were significantly higher in TGF-β2+ OMS group than normal control group and TGF-β2 group, and the late apoptosis rate in TGF-β2+ OMS group was significantly higher than that of normal control group (all at P<0.05). There were statistically significant differences in cleaved caspase-9/procaspase-9, bax, and bax/bcl-2 among the four groups ( F=4.40, 7.98, 4.61; all at P<0.05). The bax/bcl-2 expression was significantly increased in TGF-β2+ OMS group in comparison with normal control group, and the expressions of cleaved caspase-9/procaspase-9, bax, and bax/bcl-2 were significantly elevated in TGF-β2+ OMS group compared with TGF-β2 group (all at P<0.05). LDH activity in the normal control group, TGF-β2 group, TGF-β2+ OMS group and OMS group was (783.99±79.97), (913.16±196.86), (2 529.06±240.21), and (2 134.29±138.96) μmol/(min·L), respectively, showing a statistically significant difference ( F=24.95, P<0.05). Compared with normal control group and TGF-β2 group, LDH activity in TGF-β2+ OMS group was increased, and the differences were statistically significant (both at P<0.05). SOD activity in the normal control group, TGF-β2 group, TGF-β2+ OMS group and OMS group was (50.35±0.97), (41.61±4.56), (28.88±3.26), and (37.61±4.83) μmol/(min·L), respectively, showing a statistically significant difference ( F=5.71, P<0.05). SOD activity was reduced in TGF-β2+ OMS group compared with normal control group and TGF-β2 group, reduced in OMS group compared with normal control group, and the differences were statistically significant (all at P<0.05). Conclusions:AGTR1 blocker OMS can promote the apoptosis of HTF effectively.Mitochondrial apoptosis pathway mediated by bax/bcl-2/caspase-9 and oxidative stress pathway are the potential mechanisms that OMS regulates the apoptosis of HTF.
3.Study on predicting the risk of retinal vein occlusion based on nomogram model and systemic risk factors
Meilin SHAO ; Meimei REN ; Wenyi ZHANG ; Zhuoyan YANG ; Yidan WU ; Jianming WANG ; Lijun WANG
Chinese Journal of Ocular Fundus Diseases 2023;39(5):381-386
Objective:To establish and preliminarily validate a nomogram model for predicting the risk of retinal vein occlusion (RVO).Methods:A retrospective clinical study. A total of 162 patients with RVO (RVO group) diagnosed by ophthalmology examination in The Second Affiliated Hospital of Xi'an Jiaotong University from January 2017 to April 2022 and 162 patients with age-related cataract (nRVO group) were selected as the modeling set. A total of 45 patients with branch RVO, 45 patients with central RVO and 45 patients with age-related cataract admitted to Xi'an Fourth Hospital from January 2022 to February 2023 were used as the validation set. There was no significant difference in gender composition ratio ( χ2=2.433) and age ( Z=1.006) between RVO group and nRVO group ( P=0.120, 0.320). Age, gender, blood routine (white blood cell count, hemoglobin concentration, platelet count, neutrophil count, monocyte count, lymphocyte count, erythrocyte volume, mean platelet volume, platelet volume distribution width), and four items of thrombin (prothrombin time, activated partial thrombin time, fibrinogen, and thrombin time) were collected in detail ), uric acid, blood lipids (total cholesterol, triglyceride, high-density lipoprotein, low-density lipoprotein, lipoprotein a), hypertension, diabetes mellitus, coronary heart disease, and cerebral infarction. Neutrophil/lymphocyte ratio and platelet/lymphocyte ratio were calculated. The single logistic regression was used to analyze the clinical parameters of the two groups of patients in the modeling set, and the stepwise regression method was used to screen the variables, and the column graph for predicting the risk of RVO was constructed. The Bootstrap method was used to repeated sample 1 000 times for internal and external verification. The H-L goodness-of-fit test and receiver operating characteristic (ROC) curve were used to evaluate the calibration and discrimination of the nomogram model. Results:After univariate logistic regression and stepwise regression analysis, high density lipoprotein, neutrophil count and hypertension were included in the final prediction model to construct the nomogram. The χ2 values of the H-L goodness-of-fit test of the modeling set and the validation set were 0.711 and 4.230, respectively, and the P values were 0.701 and 0.121, respectively, indicating that the nomogram model had good prediction accuracy. The area under the ROC curve of the nomogram model for predicting the occurrence of post-stroke depression in the modeling set and the verification set was 0.741 [95% confidence interval ( CI) 0.688-0.795] and 0.741 (95% CI 0.646-0.836), suggesting that the nomogram model had a good discrimination. Conclusions:Low high density lipoprotein level, high neutrophil count and hypertension are independent risk factors for RVO. The nomogram model established based on the above risk factors can effectively assess and quantify the risk of post-stroke depression in patients with cerebral infarction.
4.Radiofrequency ablation versus partial nephrectomy for the treatment of clinical stage 1 renal masses: a systematic review and meta-analysis.
Shangqian WANG ; Chao QIN ; Zhihang PENG ; Qiang CAO ; Pu LI ; Pengfei SHAO ; Xiaobing JU ; Xiaoxin MENG ; Qiang LU ; Jie LI ; Meilin WANG ; Zhengdong ZHANG ; Min GU ; Wei ZHANG ; Changjun YIN
Chinese Medical Journal 2014;127(13):2497-2503
BACKGROUNDOver the past two decades, the clinical presentation of renal masses has evolved, where the rising incidence of small renal masses (SRMs) and concomitant minimal invasive treatments have led to noteworthy changes in paradigm of kidney cancer. This study was to perform a proportional meta-analysis of observational studies on perioperative complications and oncological outcomes of partial nephrectomy (PN) and radiofrequency ablation (RFA).
METHODSThe US National Library of Medicine's life science database (Medline) and the Web of Science were exhaustly searched before August 1, 2013. Clinical stage 1 SRMs that were treated with PN or RFA were included, and perioperative complications and oncological outcomes of a total of 9 565 patients were analyzed.
RESULTSPatients who underwent RFA were significantly older (P < 0.001). In the subanalysis of stage T1 tumors, the major complication rate of PN was greater than that of RFA (laparoscopic partial nephrectomy (LPN)/robotic partial nephrectomy (RPN): 7.2%, open partial nephrectomy (OPN): 7.9%, RFA: 3.1%, both P < 0.001). Minor complications occurred more frequently after RFA (RFA: 13.8%, LPN/RPN: 7.5%, OPN: 9.5%, both P < 0.001). By multivariate analysis, the relative risks for minor complications of RFA, compared with LPN and OPN, were 1.7-fold and 1.5-fold greater (both P < 0.01), respectively. Patients treated with RFA had a greater local progression rate than those treated by PN (RFA: 4.6%, LPN/RPN: 1.2%, OPN: 1.9%, both P < 0.001). By multivariate analysis, the local tumor progression for RFA versus LPN/RPN and OPN were 4.5-fold and 3.1-fold greater, respectively (both P < 0.001).
CONCLUSIONSThe current data illustrate that both PN and RFA are viable strategies for the treatment of SRMs. Compared with PN, RFA showed a greater risk of local tumor progression but a lower major complication rate, which is considered better for poor candidates. PN is with no doubt the golden treatment for SRMs, and LPN has been widely accepted as the first option for nephron-sparing surgery by experienced urologists. RFA may be the best option for select patients with significant comorbidity.
Catheter Ablation ; adverse effects ; methods ; Humans ; Kidney Neoplasms ; surgery ; therapy ; Nephrectomy ; adverse effects ; methods