1.Expression of anoctamin 1 in Chinese hamster ovary cells and properties of its ion channel
Feng HAO ; Hao WANG ; Huijing XU ; Qing FANG ; Li ZHANG ; Hangfei ZHU ; Yunqiao ZHANG ; Xiaohong JU
Journal of Jilin University(Medicine Edition) 2014;(6):1226-1231
Objective To investigate the expression of anoctamin 1 in Chinese hamster ovary cells (CHO)and to analyze the functional properties of its ion channel,and to provide experimental basis for study on the physiological function of calcium-activated chloride channel.Methods The whole sequence of anoctamin 1 was obtained by PCR technique and was subcloned into pcDNA3.1 to construct the expression vector pcDNA3.1-anoctamin 1 was transfected into CHO by liposome-mediated transfection and the CHO stably expressing anoctamin 1 were aquired by selection with zeocin. The expression and distribution of anoctamin 1 in CHO were measured by RT-PCR technique and inverted fluorescence microscope.The functional properties of anoctamin 1 were measured with halide-sensitive fluorescence proteins YFP-H148Q/I152L.The PBS buffer solution with calcimycin and high concentration of iodine ion was used as experimental group,andthe PBS buffer solution without calcimycin and high concentration of iodine ion was used as control group.Results The results of digestion and sequencing confirmed that anoctamin 1 was cloned into pcDNA3.1 by electrophoresis and blast. The results of RT-PCR and inverted fluorescence microscope indicated that anoctamin 1 was expressed in CHO. The results of I- influx as measured by halide-sensitive fluorescence proteins YFP-H148Q/I152L showed that anoctamin 1 had the more functional properties of trans-epithelial transporting I-,and the transporting speed in experimental group was higher than that in control group (P<0.05).Conclusion Anoctamin 1 can be expressed highly in the CHO;Anoctamin 1 expressed in CHO has the characteristics of calcium-activated chloride channel.
2.Diagnosis and treatment of pre-hepatic (sinusoidal) non-cirrhotic portal hypertension and related difficulties
Journal of Clinical Hepatology 2019;35(1):13-17
Pre-hepatic (sinusoidal) non-cirrhotic portal hypertension is a group of vascular heterogeneous diseases with portal hypertension as the prominent manifestation and has a complex etiology. Compared with the patients with cirrhotic portal hypertension, the patients with pre-hepatic (sinusoidal) non-cirrhotic portal hypertension have normal liver functions, normal or a mild increase in hepatic venous pressure gradient, and better prognosis. Pre-hepatic (sinusoidal) non-cirrhotic portal hypertension may easily be misdiagnosed as unexplained cirrhotic portal hypertension, and liver pathology is the gold standard for diagnosis. Non-selective β-receptor blockers and endoscopy are major therapies for this disease, but there is a lack of high-quality clinical evidence from large-scale prospective multicenter studies.
3.Comparative Study on Electroacupuncture Versus Warm Needling for Low Back Pain
Wenshun LI ; Xuying YUE ; Jinhai XU ; Yinghao ZHOU ; Shuyan SUN ; Hangfei ZHANG ; Litao YANG ; Lin ZHUANG ; Wen MO
Shanghai Journal of Acupuncture and Moxibustion 2016;35(7):866-869
Objective To observe and compare the clinical efficacies between electroacupuncture and warm needling in treating low back pain.Method Seventy-eight eligible low back pain patients were randomized into group A of 28 cases, group B of 26 cases, and group C of 24 cases. Group A was intervened by electroacupuncture, group B was by warm needling, and group C was by medication. The short-form McGill Pain Questionnaire, Japanese Orthopaedic Association Scores (JOA), and Oswestry Disability Index were observed before and after treatment, and the therapeutic efficacies were compared.Result In group A, the McGill item scores [Sensory Pain Rating Index (S-PRI), Affective Pain Rating Index (A-PRI)] respectively after 1-week and 2-week treatment as well as in the 1-month and 3-month follow-up were significantly different from that before treatment (P<0.01,P<0.05). In group B and C, the McGill item scores after 2-week treatment and in the 1-month and 3-month follow-up were significantly different from that before treatment in the same group (P<0.01,P<0.05). The JOA and Oswestry scores were significantly changed respectively after 1-week and 2-week treatment and in the 1-month and 3-month follow-up in the three groups compared with that before treatment (P<0.05,P<0.01). After 1-week and 2-week treatment and in the 1-month and 3-month follow-up, the JOA and Oswestry scores in group A were significantly different from that in group C (P<0.05,P<0.01). In the 1-month and 3-month follow-up, the JOA scores in group B were significantly different from that in group C (P<0.05). The total effective rate was 85.7% in group A and 73.1% in group B, both significantly higher than 58.3% in group C (P<0.05). Conclusion Electroacupuncture and warm needling both can produce a significant efficacy in treating low back pain, but warm needling acts comparatively slowly and is less safe.
4.Correlation of serum angiopoietin-1,angiopoietin-2,and angiopoietin-1/angiopoietin-2 ratio with HBV DNA and alanine aminotransferase in patients with chronic hepatitis B or liver cirrhosis
Minghua LIN ; Yuan CHANG ; Fang LIU ; Yanxiang HUANG ; Hangfei XU
Journal of Clinical Hepatology 2024;40(6):1126-1129
Objective To investigate the correlation of serum angiopoietin-1(Ang-1),angiopoietin-2(Ang-2),and Ang-1/Ang-2 ratio with HBA DNA and alanine aminotransferase(ALT)in patients with chronic hepatitis B(CHB)or liver cirrhosis.Methods Clinical data and serum specimens were collected from 99 patients with CHB and 59 patients with liver cirrhosis who were admitted to Beijing YouAn Hospital,Capital Medical University,from March 2018 to October 2019,and 46 individuals who underwent physical examination were enrolled as control group.PCR was used to measure serum HBV DNA level,and ELISA was used to measure the serum levels of Ang-1 and Ang-2.The serum levels of Ang-1 and Ang-2 and Ang-1/Ang-2 ratio were compared between groups.The Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between multiple groups,and the Bonferroni method was used for further comparison between two groups;the Spearman correlation analysis was used to investigate the correlation of Ang-1,Ang-2,and Ang-1/Ang-2 ratio with HBV DNA and ALT.Results Compared with the control group,the CHB group and the liver cirrhosis group had a significant reduction in the level of Ang-1(479.0 pg/mL and 208.4 pg/mL vs 671.0 pg/mL,both P<0.05),and compared with the CHB group,the liver cirrhosis group had a significant reduction in the level of Ang-1(P<0.001).Compared with the control group,the CHB group and the liver cirrhosis group had a significant increase in the level of Ang-2(286.1 pg/mL and 438.4 pg/mL vs 198.0 pg/mL,both P<0.001),and compared with the CHB group,the liver cirrhosis group had a significant increase in the level of Ang-2(P<0.001).Compared with the control group,the CHB group and the liver cirrhosis group had a significant reduction in Ang-1/Ang-2 ratio(1.6 and 0.5 vs 3.4,both P<0.001),and compared with the CHB group,the liver cirrhosis group had a significant reduction in Ang-1/Ang-2 ratio(P<0.001).The Spearman correlation analysis showed that in the CHB group,Ang-1 was negatively correlated with HBV DNA and ALT(r=-0.400 and-0.394,both P<0.001),Ang-2 was positively correlated with HBV DNA and ALT(r=0.365 and 0.351,both P<0.001),and Ang-1/Ang-2 ratio was negatively correlated with HBV DNA and ALT(r=-0.463 and-0.473,both P<0.001);in the liver cirrhosis group,Ang-1,Ang-2,and Ang-1/Ang-2 ratio had no correlation with HBV DNA or ALT(all P>0.05).Conclusion There are significant changes in the serum levels of Ang-1 and Ang-2 and Ang-1/Ang-2 ratio in patients with CHB or liver cirrhosis,and Ang-1,Ang-2,and Ang-1/Ang-2 ratio reflects the degree of liver injury in patients with CHB to a certain extent.
5.Noninvasive assessment of the risk of esophageal variceal bleeding from noncirrhotic portal hypertension
Hangfei XU ; Yu WANG ; Fuliang HE ; Zhenhua FAN ; Hui LIU ; Yongping YANG ; Jidong JIA ; Fuquan LIU ; Huiguo DING
Chinese Journal of Hepatology 2022;30(10):1092-1099
Objective:To verify Baveno VI criteria, Expanded-Baveno VI criteria, liver stiffness×spleen diameter-to-platelet ratio risk score (LSPS), and platelet count/spleen diameter ratio (PSR) in evaluating the severity value of esophageal varices (EV) in patients with non-cirrhotic portal hypertension (NCPH).Methods:111 cases of NCPH and 204 cases of hepatitis B cirrhosis who met the diagnostic criteria were included in the study. NCPH included 70 cases of idiopathic non-cirrhotic portal hypertension (INCPH) and 41 cases of nontumoral portal vein thrombosis (PVT). According to the severity of EV on endoscopy, they were divided into the low-bleeding-risk group (no/mild EV) and the high-bleeding-risk group (moderate/severe EV). The diagnostic value of Baveno VI and Expanded-Baveno VI criteria was verified to evaluate the value of LSPS and PSR for EV bleeding risk severity in NCPH patients. The t-test or Mann-Whitney U test was used to compare the measurement data between groups. Comparisons between counting data groups were performed using either the χ2 test or the Fisher exact probability method. Results:Considering endoscopy was the gold standard for diagnosis, the missed diagnosis rates of low/high bleeding risk EVs in INCPH/PVT patients with Baveno VI and Expanded-Baveno VI criteria were 50.0%/30.0% and 53.8%/50.0%, respectively. There were no statistically significant differences in platelet count (PLT), spleen diameter, liver stiffness (LSM), LSPS, and PSR between low-bleeding-risk and high-bleeding-risk groups in INCPH patients, and the area under the receiver operating characteristic curve (AUC) of LSPS and PSR was 0.564 and 0.592, respectively ( P=0.372 and 0.202, respectively). There were statistically significant differences in PLT, spleen diameter, LSPS, and PSR between the low and high-bleeding risk groups in PVT patients, and the AUCs of LSPS and PSR were 0.796 and 0.833 ( P=0.003 and 0.001, respectively). In patients with hepatitis B cirrhosis, the Baveno VI and Expanded-Baveno VI criteria were used to verify the low bleeding risk EV, and the missed diagnosis rates were 0 and 5.4%, respectively. There were statistically significant differences in PLT, spleen diameter, LSM, LSPS and PSR between the low-bleeding-risk and high-bleeding-risk groups ( P<0.001). LSPS and PSR AUC were 0.867 and 0.789, respectively ( P<0.05). Conclusion:Baveno VI and Expanded-Baveno VI criteria have a high missed diagnosis rate for EVs with low bleeding risk in patients with INPCH and PVT, while LSPS and PSR have certain value in evaluating EV bleeding risk in PVT patients, which requires further clinical research.
6.Association between the rs1800591 variation of the microsomal triglyceride transfer protein gene and the risk of nonalcoholic fatty liver disease in the elderly population
Jinhan ZHAO ; Jing ZHANG ; Yang ZHANG ; Xiaoyi XU ; Yusong GOU ; Hangfei XU ; Yan WAN ; Jian WU
Journal of Clinical Hepatology 2023;39(3):552-561
Objective To investigate the association between the polymorphism of the microsomal triglyceride transport protein (MTTP) gene at rs1800591 locus and the risk of nonalcoholic fatty liver disease (NAFLD) in the elderly population. Methods The clinical cohort of this study was established in Menkuang Hospital, Beijing Jingmei Group General Hospital. A total of 1098 healthy elderly volunteers were recruited for physical examination in communities in Mentougou District of Beijing, China, from January 11, 2020 to September 30, 2021, among whom there were 614 patients with NAFLD and 484 individuals without NAFLD. Gene microarray was used to determine the genotypes of MTTP rs1800591; demographic data were collected, and blood biochemical parameters were measured. The independent samples t -test was used for comparison of normally distributed continuous data between groups, and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between groups; the chi-square test was used for comparison of categorical data between groups. The chi-square test was used to investigate whether the distribution of genotype frequency was in accordance with Hardy-Weinberg equilibrium. The unconditional logistic regression model was used to calculate odds ratio ( OR ) and its 95% confidence interval ( CI ) to investigate the association of gene polymorphism with the risk of NAFLD and other comorbidities. Results There were significant differences in sex and age between the two groups ( P < 0.05). Compared with the non-NAFLD group, the NAFLD group had significantly higher levels of body mass index (BMI), waist-hip ratio, triglyceride, alanine aminotransferase, aspartate aminotransferase, controlled attenuation parameter (CAP), and liver stiffness measurement and a significantly lower level of high-density lipoprotein (HDL) (all P < 0.05). Compared with the non-NAFLD group, the NAFLD group had a significantly higher proportion of patients with hypertension, diabetes, obesity, and metabolic syndrome (all P < 0.05). The distribution of genotype frequency at MTTP rs1800591 locus was in accordance with Hardy-Weinberg equilibrium in the control group ( χ 2 =1.097, P =0.29). There were a significant differences in the genotype and the distribution of alleles at MTTP rs1800591 locus between the patients with NAFLD and the control group (all P < 0.001). In the total population, there was a significantly lower carrying rate of T allele (GT+TT, n =351) in male individuals, and the individuals carrying T allele had significantly higher BMI and CAP than those carrying GG allele ( n =747) ( P < 0.001). Compared with the individuals who did not carry T allele, the individuals carrying T allele (GT+TT, n =232) had a significantly higher proportion of patients with obesity and a significantly lower NFS score ( P < 0.05). As for the individuals with NAFLD, the individuals carrying T allele had a significantly lower proportion of male individuals, a significantly lower waist-hip ratio, and a significantly higher level of HDL compared with those who did not carry T allele (GG, n =382), and the GT+TT group had a significantly lower NFS score than the GG group (all P < 0.05). The non-conditional logistic regression analysis showed that after adjustment for the confounding factors of sex, age, and BMI, the GT+TT genotype at MTTP rs1800591 locus significantly increased the risk of NAFLD ( OR =1.643, 95% CI : 1.226-2.203, P =0.001), and carrying T allele also increased the risk of obesity in the total population ( OR =1.371, 95% CI : 1.051-1.788, P =0.02). Conclusion MTTP rs1800591 polymorphism is associated with the development of NAFLD in the elderly population, and carrying T allele may promote hepatic steatosis and increase the risk of obesity in NAFLD, while it may inhibit the progression of liver fibrosis.