BACKGROUND: Autophagy, oxidative stress and inflammatory reactions play an important role in non-alcoholic fatty liver disease. Curcumin has biological activities such as regulating autophagy, oxidative stress and inflammatory reaction. OBJECTIVE: To explore the effect and underlying mechanism of curcumin on experimental non-alcoholic fatty liver disease rats. METHODS: Non-alcoholic fatty liver model was established in rats fed 8-week high-fat diets. Forty healthy SPF male Sprague-Dawley rats were randomly divided into control group, model group, curcumin treatment group, and curcumin and 3-methyladenine (3-MA, an autophagic flux blocker) treatment group. At the end of 8 weeks of high-fat diet, control and model groups were given PBS intragastrically, curcumin treatment group given curcumin 500 mg/kg per day intragastrically, and curcumin+3-MA given curcumin 500 mg/kg per day intragastrically and 3-MA 2 mg/kg per day intraperitoneally. The interventions in each group were given for 8 continuous weeks. The biochemical parameters including serum alanine aminotransferase, aspartate aminotransferase, total triglyceride, total cholesterol, fasting blood glucose level were measured in rats. Oil red O staining was used to characterize the change of hepatic pathology. The ultrastructure of mitochondria was examined by transmission electron microscopy. The hepatic malondialdehyde level and superoxide dismutase activity were measured by thiobarbituric acid method and xanthine oxidase method, respectively. Western blot assay was performed to detect the expression level of autophagic molecular signals including P62, Beclin, LC3B and nuclear factor-KB. The experimental protocol was approved by the Animal Ethic Committee of Renmin Hospital of Wuhan University (approval No. 2018-541). RESULTS AND CONCLUSION: The serum alanine aminotransferase, aspartate aminotransferase, total triglyceride, and total cholesterol levels were significantly higher in the model group than the control group (P < 0.05). Compared with the model group, curcumin significantly decreased the levels of serum alanine aminotransferase, aspartate aminotransferase, total triglyceride, and total cholesterol (P < 0.05), but this effect was partly inhibited by 3-MA (P < 0.05). There was more cellular lipid deposition in the model group than the control group. Compared with the model group, curcumin significantly decreased cellular lipid deposition, but the decrease was partly inhibited by 3-MA. Compared with the control group, mitochondrial edema and cristae rupture (or even completely disappearing) were easily found in the model group. Curcumin significantly attenuated mitochondrial injury, which was partly inhibited by 3-MA. Hepatic superoxide dismutase activity in the model group was significantly lower than that in control group, and it significantly increased after curcumin treatment. The hepatic superoxide dismutase activity in the curcumin+3-MA group was higher than that in model group but lower than that in the curcumin group. Hepatic malondialdehyde level in the model group was higher than that in the control group, and it significantly decreased after curcumin treatment. Whereas the hepatic malondialdehyde level in the curcumin+3-MA group was lower than that in model group but higher than that in the curcumin group. Compared with the control group, the model group showed significantly increased expressions of P62, nuclear factor-KB but decreased expressions of Beclin-1 and LC3II/LC3I (all P < 0.05). Curcumin significantly decreased the expressions of P62 and nuclear factor -kB and increased the expressions of Beclin-1 and LC3II/LC3I; however, these changes were partly inhibited by 3-MA (all P < 0.05). Therefore, curcumin can effectively prevent hepatic steatosis in experimental non-alcoholic fatty liver disease rats by regulating inflammatory reaction and oxidative stress via activation of autophagy.

Objective To investigate the risk factors of electrocoagulation syndrome after endoscopic submucosal dissection (ESD) in patients with colorectal lesions.Methods Clinical data of 145 patients with colorectal mucosal lesions undergoing ESD in People's Hospital of Wuhan University between September 2010 and September 2015 were retrospectively studied.Results Among 45 patients,post endoscopic submucosal dissection electrocoagulation syndrome (PEECS) was developed in 32 cases (22％).The median age in PEECS group was higher (t =-5.783,P =0.000),the median lesion size was larger(t =-5.590,P =0.000),the median length of hospital stay was longer (t =-6.841,P =0.000) than those in non-PEECS group.Univariate regression analysis showed PEECS was associated with the age,lesion size,lesion location,length of hospital stay,malignant tumor,polyps type,resection modality.Multivariable logistic regression analysis showed that the independent risk factors for the development of electrocoagulation syndrome were age ＞65 year (OR =1.123,95％ CI:1.013-1.244,P =0.027),lesion size ＞ 3.5 cm (OR =1.173,95％ CI:1.015-1.357,P =0.031),malignant tumor (OR =3.498,95 ％ CI:1.460-8.379,P =0.005),hospital stay ＞ 10 d (OR =2.480,95％ CI:1.346-4.569,P =0.004),non-rectal lesions (OR =12.612,95％ CI:3.446-46.157,P =0.000).Conclusion Attention should be paid for colorectal lesion patients with high risk of PEECS,when endoscopic submucosal dissection is performed.

Objective To explore the characteristics of intestinal Microbiota in T2DM patients by two molecular fingerprint technologies,and investigate the correlation of intestinal microbiota and T2DM,and evaluate the application value of two fin-gerprint technologies.Methods Fecal samples of 8 healthy groups and 7 diabetes patients were collected.Then the total DNA of gut microbiota was extracted.Through the analysis of products by two molecular fingerprints of ERIC-PCR and DGGE-PCR,ecological characteristics of diversity and similarity of gut microbiota were obtained in healthy groups and dia-betes patients.Results Compared to healthy groups,the number of bands and Shannon-Wiener index of gut microbiota in di-abetes patients was decreased but no statistical significance.The similarity in patients group was declining(P <0.05),and the construction of gut microbiota was inclined to differ.Two fingerprint technologies of ERIC and DGGE could directly re-flect the diversity of gut microbiota and were the modern molecular biological techniques without depending on cultivation. ERIC was simple and convenient,had a better reflection of microbial diversity,but gel band cutting and regarded asa proper approach with higher diffraction efficiency and excellent repetition to studysequencing couldn’t be performed since there were more influencing factors on the experiment.DGGE could better reflect the ecological characteristics such as microbial diversity and similarity,and selecting bands,gel band cutting and sequencing could be done.Conclusion The composition and construction of gut microbiota in diabetes patients were changed,which suggests the occurrence of the disease had the correlation with gut microbiota.ERIC and DGGE is regarded as a proper approach with higher diffraction efficiency and ex-cellent repetition to study intestinal microbiota,but also gel band cutting,sequencing,bacteria identification can be performed by DGGE,both can be used in combination.

Objective To evaluate the change of serum creatinine (Scr) before and after administration of contrast agent in different dose,to observe the difference of dog's kidney tissue with electron microscopy and investigate the effect of contrast agent on renal function.Methods Twelve dogs were divided into four groups randomly:the control group,the low dose group,the moderate dose group and high dose group.After the administration of different doses of iodine contrast agent at the same rate,the changes of Scr and microscopic structure were compared before administration and 48 hours later.Results The differences of Scr before and 48 hours after administration were (4.6±1.6) μmol/L,(6.7±2.5) μmol/L,(6.9±4.5) μmol/L,(5.1± 1.9) μmol/L for control group,low dose group,moderate dose group and high dose group,respectively.There was no statistically significant difference among the groups (P ＞0.05).In high dose group,the mitochondria of tubular epithelial cells were swelling and obvious vacuoles were observed.Only a small amount of vacuoles existed in the renal tubular epithelial cells in low dose group.Conclusion Compared with the moderate and high dose group,the low-dose iodine contrast agent have less damage to the kidney cells of the dogs.

Objective:To explore the effect of mirror visual feedback combined with exercise relearning in rehabilitation of upper extremity dysfunction after stroke.Methods:Patients with upper extremity dysfunction received stroke rehabilitation treatment in the Nanyang No. 9 People's Hospital from August 2016 to August 2018 were enrolled prospectively. According to the random number table method, they were divided into either mirror visual feedback combined with exercise relearning group (mirror therapy group) or exercise relearning alone group (control group). The rehabilitation treatment was 1-2 times a day, 5 d a week, and 1 week was a course of treatment, for a total of 4 courses. Fugl-Meyer Assessment (FMA) was used to assess upper extremity function. Carroll Upper Extremity Function Test (UEFT) was used to assess hand function. Action Research Arm Test (ARAT) was used to assess upper extremity operability and flexibility. Motor Assessment Scale (MAS) was used to assess the motor function of hands, fingers and upper extremities. Visual Analogue Scale (VAS) was used to assess upper extremities pain. The modified Barthel Index (MBI) was used to assess self-care ability in daily life. The modified Ashworth Spasm Scale was used to assess the degree of spasm of shoulder joints, wrist joints, and elbow joints. According to the FMA score, the clinical efficacy of both groups of patients was evaluated, and a score of >31 was defined as excellent.Results:A total of 60 stroke patients with upper extremity dysfunction were enrolled ( n=30 in the mirror therapy group and n=30 in the control group). There were no statistical differences between the two groups in age, gender, course of disease, stroke type, and stroke location, as well as each baseline score. After treatment, the FMA score, UEFT score, ARAT score, MAS score, MBI score and the proportion of the modified Ashworth grade Ⅰ and grade Ⅰ + were significantly higher than those before treatment in both groups (all P<0.05), while VAS score and the proportion of modified Ashworth grade Ⅲ and grade Ⅳ was significantly lower than those before treatment (all P<0.05). After treatment, FMA score, UEFT score, ARAT score, MAS score, MBI score, as well as the proportion of Ashworth grade Ⅰ and grade Ⅰ + in the mirror therapy group was significantly higher than that in the control group (all P<0.05), while VAS score and the proportion of modified Ashworth grade Ⅲ were significantly lower than those in the control group ( P<0.05). According to the FMA score, the excellent and good rate of treatment in the mirror therapy group was significantly higher than that in the control group (93.3% vs. 70.0%; χ2=5.455, P=0.020). Conclusions:The curative effect of mirror visual feedback combined with exercise relearning is superior to exercise relearning alone in rehabilitation therapy for upper extremity dysfunction after stroke.

Objective To explore the association between patatin-like phospholipase domaincontaining protein 3 (PNPLA3) gene rs738409 polymorphism and the susceptibility of non-alcoholic fatty liver disease (NAFLD).Methods Data bases were comprehensively searched to retrace all the related studies on the association between PNPLA3 gene rs738409 polymorphism and susceptibility.Of NAFLD,the pooled OR with 95％ CI of the association between PNPLA3 gene rs738409 polymorphism and NAFLD susceptibility were performed using different genetic models.Subgroup analysis based on the source of population and sensitivity analysis was performed to detect the stability of results.Results 28 original studies with 6 216 patients and 8 218 controls were involved in the final combination of data.Findings from the meta-analyses showed that there were strong associations between PNPLA3 gene rs738409 polymorphism and the susceptibility of NAFLD,under different genetic model comparisons [GG vs.CC:OR=2.42,95％CI:1.83-3.21,P＜0.001 ;CG vs.CC:OR=1.28,95％CI:1.15-1.43,P＜0.001 ; CG+GG vs.CC:OR=1.31,95％CI:1.17-1.46,P＜ 0.001 ; GG vs.CC + GC:OR=2.26,95％CI:1.76-2.90,P＜0.001].Similar results were found in both Asian and Caucasian populations.Conclusion Results from the Meta-analysis strongly suggested that there appeared significant association between PNPLA3 gene rs738409 polymorphism and the susceptibility of NAFLD.

Objective To study whether matrix metalloproteinases-9 (MMP)-1562C/T (rs3918242) and MMP-2-1306C/T (rs243865) were associated with the susceptibility on nonalcoholic fatty liver disease (NAFLD) and the interactions between the two factors and central obesity.Methods Genotypes of 545 patients and 636 subjects with NAFLD under control were examined by polymerase chain reaction-based restriction fragment length polymorphism (PCR-RFLP).Unconditional logistic regression (ULR) was performed to assess the NAFLD risk.The geneenvironment interactions on the risk of NAFLD were explored by generalized multifactor dimensionality reduction (GMDR) and ULR methods.Results Results from the case-control analysis indicated that there was an increased risk of developing NAFLD for MMP-9 rs3918242 (TT/CT) genotype carriers,when compared with the non-carriers (CC),with OR=1.67,95％ CI:1.32-2.12,P=0.001;Adjusted OR=1.65,95％CI:1.31-2.01 (P=0.008).However,risk reduction of NAFLD was found when MMP-2 rs243865 (TT/CT) genotype carriers compared with the non-carriers (CC),with OR=0.68,95％CI:0.53-0.86,P=0.001;with adjusted OR=0.66,95％CI:0.49-0.90 (P=0.007).Data from the GMDR showed that gene-environment interaction among rs3918242 and central obesity on the risk of NAFLD might be significant (P=0.001).By using the ULR method,subjects as central obesity-positive but with genotype CT/TT,appeared having 4.50 (95％ CI:2.78-7.17,P =0.007) times risk of NAFLD,when compared to the central obesity-negative subjects with genotype CC after adjusting for the covariates.Conclusion MMP-9 rs3918242,MMP-2 rs243865 were associated with risk of NAFLD while both rs3918242 and central obesity showing synergistic effects on the risk of the NAFLD.

Objective:To investigate the short- and long-term outcomes of fetuses with selective fetal growth restriction (sFGR).Methods:A retrospective study was conducted on monochorionic diamniotic (MCDA) twins with sFGR admitted to the Neonatal Intensive Care Unit of Peking University Third Hospital from September 2017 to December 2019. MCDA neonates delivered during the same period without significant complications were selected as the control group. MCDA twins with sFGR were divided into type Ⅰ, Ⅱ, and Ⅲ groups and then further divided into the larger and the smaller fetus subgroups according to the birth weight. These children were followed up by telephone at 2-3 years old. Height-for-age and weight-for-age Z-scores were calculated. Ages and Stages Questionnaire-Third Edition (ASQ-3) was used to determine comprehensive development. Independent sample t-test, one-way analysis of variance, non-parameter test, and Chi-square test (or rank-sum test) were used for statistical analysis. Results:(1) A total of 116 pregnant women with sFGR (232 neonates) were enrolled in this study. There were 43, 40, and 33 mothers and 86, 80, and 66 newborns in type Ⅰ, Ⅱ, and Ⅲ groups, respectively. The control group included 31 pregnant women and 62 neonates. The gestational age at onset of sFGR was younger in the type Ⅱ and Ⅲ groups than in type Ⅰ group [(23.8±4.8) and (24.1±3.1) vs (27.0±6.1) weeks, F=5.19, P<0.05; all P<0.017 during pairwise comparisons]. (2) The incidence of sepsis and treatment abandonment/death in neonates in type Ⅱ and Ⅲ groups were higher than those in type Ⅰ and control groups [neonatal sepsis: 11.3% (9/80) and 6.1% (4/66) vs 2.3% (2/86) and 0.0% (0/62), χ2=6.30, P=0.001; death or treatment abandonment rate:13.8% (11/80) and 10.6% (7/66) vs 3.5% (3/86) and 0.0% (0/62), χ2=4.68, P=0.003; all P<0.017 during pairwise comparisons]. In cases with type Ⅱ or type Ⅲ sFGR, the risk of digestive system diseases was significantly higher in the smaller fetus group than in the larger fetus group [type Ⅱ: 46.2% (37/80) vs 38.7% (31/80), χ2=16.72; type Ⅲ: 47.0% (31/66) vs 34.8% (23/66), χ2=39.69; both P<0.001], while the rate of respiratory system diseases was lower in the smaller fetus group [type Ⅱ: 35.0% (28/80) vs 45.0% (36/80), χ2=36.85; type Ⅲ: 37.9% (25/66) vs 45.4% (30/66), χ2=12.55; both P<0.001]. The incidence of neonatal sepsis in smaller fetuses was higher than that in larger ones in type Ⅱ sFGR [7.5% (6/80) vs 3.7% (3/80), χ2=4.68, P=0.034]. The incidence of neurological complications in larger fetuses was higher than that in smaller ones in type Ⅲ sFGR [15.1% (10/66) vs 4.5% (3/66), χ2=5.72, P<0.001]. (3) In type Ⅱ group, seven neonates died (one case of cerebral hemorrhage, two cases of gastrointestinal perforation, two cases of septic shock, and two cases of necrotizing enterocolitis), and four cases withdrew the treatment. In type Ⅲ group, four neonates died (two cases of necrotizing enterocolitis, one case of gastrointestinal perforation, and one case of cerebral hemorrhage), and three cases withdrew from the treatment. (4) Totally, 71 children in type Ⅰ, 61 in type Ⅱ, and 58 in type Ⅲ group were followed up at the age of 2-3. Children with type Ⅱ or type Ⅲ sFGR lagged behind those in type Ⅰ group and control group in physical growth [ M ( P25- P75), Z-scores:－0.46 (－0.87-0.42),－0.35 (－0.62-0.71), 0.05 (－0.61-0.51), and 0.14 (－0.57-0.75); H=6.20, P=0.001]. In type Ⅱ and Ⅲ groups, the smaller fetuses lagged the larger fetuses in physical growth at 2-3 years of age. ASQ-3 scores in communication, gross motor, fine motor, problem-solving and personal-social areas were all lower in type Ⅱ and Ⅲ groups than in type Ⅰ and control groups. ASQ-3 scores in the five dimensions of the smaller fetuses in the type Ⅱ group were lower than those of the larger fetuses. In the type Ⅲ group, the smaller fetuses had lower ASQ-3 scores in communication and gross motor than the larger ones [communication ability: (42.6±18.8) vs (56.4±9.4) scores, t=19.63, P<0.001; gross motor: (45.5±19.7) vs (54.5±9.7) scores, t=12.64, P=0.003]. Conclusion:The neonatal morbidity is significantly increased in type Ⅱ and Ⅲ sFGR, and babies lagged others in height, weight, and ASQ-3 score at 2-3, which is worthy of early attention.

Objective:To clarify the infection and epidemic characteristics of the human metapneumovirus (HMPV) in Chinese patients with febrile respiratory syndrome (FRS), and to provide important baseline data for clinical diagnosis, treatment, prevention and control of HMPV-induced respiratory tract diseases in China.Methods:FRS cases from January 2009 to June 2021 in 9 provinces in China, including Beijing, Hebei, Jilin, Shandong, Shaanxi, Xinjiang, Anhui, Guangdong, Hunan were retrospectively analyzed for their respiratory samples, clinical and epidemic data.The respiratory samples were detected for HMPV by quantitative real-time PCR.Results:A total of 11 660 cases were tested for HMPV, involving 296 (2.54%) HMPV-positive cases.Among 296 HMPV-positive cases, 218 were single HMPV infection, and 78/296 (26.35%) were co-infected with one or more respiratory viruses.HMPV mainly affected children under 5 years of age (3.10%), and in this population, the proportion of pneumonia in HMPV co-infection cases was significantly higher than that of single HMPV infection.HMPV could be detected all year round, which was more popular in winter and spring, with the peak of HMPV epidemic in March.Conclusions:HMPV is one of the important pathogens causing acute respiratory infection in children, showing a clear seasonal epidemic.HMPV can be infected alone or in combination with other respiratory viruses, which may increase the risk of pneumonia in children.

Gene editing technology has been a hotspot in the field of biotechnology. CRISPR/Cas systems are efficient gene editing tools because of its specificity, simplicity and flexibility, these features enabled the rapid application of CRISPR/Cas systems in a variety of organisms. Moreover, the combination of transcriptional activator with dead Cas protein can achieve specific regulation of gene expression at the transcription level, which has made important contributions to the development of biotechnology in medical and agriculture. Overexpression of foreign genes is a common method to verify gene function and regulation. However, due to the limitation of vector capacity, it is difficult to achieve overexpression of multiple genes. CRISPR/Cas9 activation system can regulate the expression of multiple genes under the guidance of different guide RNAs to verify gene functions at the regulatory level. This review summarizes the composition of the CRISPR/Cas9 activation system and different activation strategies, and summarizes solutions for excessive activation. It may facilitate the application of CRISPR/Cas9 activation system in genetic improvement of cotton and herbicide resistance research.
Biotechnology ; CRISPR-Cas Systems/genetics* ; Gene Editing ; Phenotype ; RNA, Guide, Kinetoplastida/metabolism*