Traditional Chinese herbal extracts inducing autophagy as a novel approach in therapy of nonalcoholic fatty liver disease

Non-alcoholic fatty liver disease(NAFLD) is one of the leading causes of chronic liver diseases around the world due to the modern sedentary and food-abundant lifestyle, which is characterized by excessive fat accumulation in the liver related with causes other than alcohol abuse. It is widely acknowledged that insulin resistance, dysfunctional lipid metabolism, endoplasmic reticulum stress, oxidative stress, inflammation, and apoptosis/necrosis may all contribute to NAFLD. Autophagy is a protective self-digestion of intracellular organelles, including lipid droplets(lipophagy), in response to stress to maintain homeostasis. Lipophagy is another pathway for lipid degradation besides lipolysis. It is reported that impaired autophagy also contributes to NAFLD. Some studies have suggested that the histological characteristics of NAFLD(steatosis, lobular inflammation, and peri-sinusoid fibrosis) might be improved by treatment with traditional Chinese herbal extracts, while autophagy may be induced. This review will provide insights into the characteristics of autophagy in NAFLD and the related role/mechanisms of autophagy induced by traditional Chinese herbal extracts such as resveratrol, Lycium barbarum polysaccharides, dioscin, bergamot polyphenol fraction, capsaicin, and garlic-derived S-allylmercaptocysteine, which may inhibit the progression of NAFLD. Regulation of autophagy/lipophagy with traditional Chinese herbal extracts may be a novel approach for treating NAFLD, and the molecular mechanisms should be elucidated further in the near future.

DLAT as a Cuproptosis Promoter and a Molecular Target of Elesclomol in Hepatocellular Carcinoma

Objective Cuproptosis is a novel cell death pathway that was newly discovered in early 2022.However,cuproptosis is still in its infancy in many respects and warrants further research in hepatocellular carcinoma(HCC).This study aimed to analyze the mechanism of cuprptosis in HCC.Methods Herein,the tumor microenvironment infiltration landscape of molecular subtypes was illustrated using GSVA,ssGSEA,TIMER,CIBERSORT,and ESTIMATE algorithms based on the expression profile of cuproptosis-related genes(CRGs)from TCGA and GEO databases.Then,the least absolute shrinkage and selection operator regression method was applied to construct a cuproptosis signature to quantify the cuproptosis profile of HCC.Further,we explored the expression of three hub CRGs in cell lines and clinical patient tissues of HCC by Western blotting,qRT-PCR and immunohistochemistry.Finally,we examined the function of dihydrolipoamide S-acetyltransferase(DLAT)in cuproptosis in HCC by loss-of-function strategy,Western blotting and CCK8 assay.Results Three distinct molecular subtypes were identified.Cluster 2 had the greatest infiltration of immune cells with best prognosis.The cuproptosis signature was indicative of tumor subtype,immunity,and prognosis for HCC,and specifically,a low cuproptosis score foreshadowed good prognosis.DLAT was highly expressed in liver cancer cell lines and HCC tissues and positively correlated with clinical stage and grade.We also found that potent copper ionophore elesclomol could induce cuproptosis in a copper-dependent manner.Selective Cu^(++)chelator ammonium tetrathiomolybdate and downregulating DLAT expression by siRNA could effectively inhibit cuproptosis.Conclusion Cuproptosis and DLAT as a promising biomarker could help to determine the prognosis of HCC and may offer novel insights for effective treatment.

MicroRNA-143 suppresses gastric cancer cell growth and induces apoptosis by targeting COX-2

AIM:To investigate the function of microRNA-143(miR-143)in gastric cancer and explore the target genes of miR-143.METHODS:A quantitative real-time reverse transcription-polymerase chain reaction(RT-PCR)analysis was performed to evaluate miR-143 expression in gastric cancer cell lines.After transfecting gastric cancer cells with miR-143-5p and miR-143-3p precursors,Alamar blue and apoptosis assays were used to measure the respective proliferation and apoptosis rates.Cyclooxygenase-2(COX-2)expression was determined by realtime RT-PCR and Western blot assays after miR-143transfection.Reporter plasmids were constructed,and a luciferase reporter assay was used to identify the miR-143 binding site on COX-2.RESULTS:Both miR-143-5p and miR-143-3p were significantly downregulated in multiple gastric cancer cell lines.Forced miR-143-5p and miR-143-3p expression in gastric cancer cells produced a profound cytotoxic effect.MiR-145-5p transfection into gastric cancer cells resulted in a greater growth inhibitory effect(61.23%±3.16%vs 46.58%±4.28%,P<0.05 in the MKN-1cell line)and a higher apoptosis rate(28.74%±1.93%vs 22.13%±3.31%,P<0.05 in the MKN-1 cell line)than miR-143-3p transfection.Further analysis indicated that COX-2 expression was potently suppressed by miR-143-5p but not by miR-143-3p.The activity of a luciferase reporter construct that contained the 3’-untranslated region(UTR)of COX-2 was downregulated by miR-143-5p(43.6%±4.86%,P<0.01)but not by miR-143-3p.A mutation in the miR-145-5p binding site completely ablated the regulatory effect on luciferase activity,which suggests that there is a direct miR-145-5p binding site in the 3’-UTR of COX-2.CONCLUSION:Both miR-143-5p and miR-143-3p function as anti-oncomirs in gastric cancer.However,miR-143-5p alone directly targets COX-2,and it exhibits a stronger tumor suppressive effect than miR-143-3p.

Evaluation of a new method for placing nasojejunal feeding tubes

AIM:To compare fluoroscopic, endoscopic and guide wire assistance with ultraslim gastroscopy for placement of nasojejunal feeding tubes. METHODS:The information regarding nasojejunal tube placement procedures was retrieved using the gastrointestinal tract database at Tongji Hospital affiliated to Tongji Medical College. Records from 81 patients who underwent nasojejunal tubes placement by different techniques between 2004 and 2011 were reviewed for procedure success and tube-related outcomes. RESULTS:Nasojejunal feeding tubes were successfully placed in 78 (96.3%) of 81 patients. The success rate by fluoroscopy was 92% (23 of 25), by endoscopic technique 96.3% (26 of 27), and by guide wire assistance (whether via transnasal or transoral insertion)100% (23/23, 6/6). The average time for successful placement was 14.9 ± 2.9 min for fluoroscopic placement, 14.8 ± 4.9 min for endoscopic placement, 11.1 ± 2.2 min for guide wire assistance with transnasal gastroscopic placement, and 14.7 ± 1.2 min for transoral gastroscopic placement. Statistically, the duration for the third method was significantly different (P < 0.05) compared with the other three methods. Transnasal placement over a guidewire was significantly faster (P < 0.05) than any of the other approaches. CONCLUSION:Guide wire assistance with transnasal insertion of nasojejunal feeding tubes represents a safe, quick and effective method for providing enteral nutrition.

Novel biosensor-based microarray assay for detecting rs8099917 and rs12979860 genotypes

AIM:To evaluate a novel biosensor-based microarray(BBM) assay for detecting rs12979860 and rs8099917 genotypes.METHODS:Four probes specific for rs8099917C/T or rs12979860G/T detection and three sets of quality control probes were designed,constructed and arrayed on an optical biosensor to develop a microarray assay.Two sets of primers were used in a one tube polymerase chain reaction(PCR) system to amplify two target fragments simultaneously.The biosensor microarray contained probes that had been sequenced to confirm that they included the rs8099917C/T or rs12979860G/T alleles of interest and could serve as the specific assay standards.In addition to rehybridization of four probes of known sequence,a total of 40 clinical samples collected from hepatitis C seropositive patients were also tested.The target fragments of all 40 samples were amplified in a 50 μL PCR system.Ten μL of each amplicon was tested by BBM assay,and another 40 μL was used for sequencing.The agreement of the results obtained by the two methods was tested statistically using the kappa coefficient.The sensitivity of the BBM assay was evaluated using serial dilutions of ten clinical blood samples containing 10 3-10 4 white cells/μL.RESULTS:As shown by polyacrylamide gel electrophoresis,two target segments of the interleukin 28Bassociated polymorphisms(SNPs) were successfully amplified in the one-tube PCR system.The lengths of the two amplified fragments were consistent with the known length of the target sequences,137 and 159 bps.After hybridization of the PCR amplicons with the probes located on the BBM array,the signals of each allele of both the rs8099917 SNPs and rs12979860 SNPs were observed simultaneously and were clearly visible by the unaided eye.The signals were distinct from each other,could be interpreted visually,and accurately recorded using an ordinary digital camera.To evaluate the specificity of the assay,both the plasmids and clinical samples were applied to the microarray.First,30 PCR amplicons of the various SNP alleles were hybridized on the BBM microarray.Full agreement between plasmids and the BBM assay was observed,with 30/30 correct matches(100%).The kappa value for the BBM assay with plasmids was 1.00(P < 0.05).For the 40 clinical blood samples,the BBM assay hybridization and direct sequencing results were compared for each amplicon.For patient blood samples,agreement was 28/28 for rs8099917T/T,9/11 for rs8099917T/G,1/1 for rs8099917G/G,24/24 for rs12979860C/C,11/14 for rs12979860C/T,and 2/2 for rs12979860T/T.Only five clinical samples of amplicon assay and direct sequencing results were discordant and heterozygotes:2/11 rs8099917T/G and 3/14 rs12979860C/T.The agreement of outcomes between BBM assay and direct sequencing for the detection of rs8099917 and rs12979860 was 95% and 92.5%,respectively;and the corresponding kappa values were 0.88 and 0.85(A kappa value > 0.75 was defined as substantial agreement).The BBM assay and sequencing had similar specificities for detection and identification of the two SNPs and their alleles.The sensitivity evaluation showed that the BBM assay could detect and identify SNP sequences present in blood samples containing as few as 10 2 white blood cells/μL.CONCLUSION:This biosensor microarray assay was highly specific,sensitive,rapid and easy to perform.It is compatible with clinical practice for detection of rs8099917 and rs12979860.

Urgent Endoscopy in Nonvariceal Upper Gastrointestinal Hemorrhage:A Retrospective Analysis

Objective:The role of urgent endoscopy in nonvariceal upper gastrointestinal hemorrhage(NVUGIH)remains controversial.We designed a retrospective study to compare the outcomes between urgent endoscopy(within 12 h)and non-urgent endoscopy for patients with NVUGIH.Methods:A total of 540 hospitalized patients with NVUGIH were included in our study.Patients who received endoscopy within 12 h or after 12 h were divided into two groups,the urgent and non-urgent endoscopy groups,respectively.The clinical outcomes including rebleeding,mortality,endoscopic re-intervention,need for emergency surgery and interventional radiotherapy were compared between the groups.Patients with Glasgow-Blatchford scores(GBS)<12 and>12 were defined as the lower-and high-risk groups,respectively,and the predictors of rebleeding and mortality in both groups were analyzed individually.Results:Patients with NVUGIH in the urgent endoscopy group had a higher rate of rebleeding(27.6%vs.16.9%,P=0.003)and blood transfusion(73.2%vs.55.5%,P<0.001)than those in the non-urgent endoscopy group,while the mortality and the length of hospitalization were not significantly different between the groups(P>0.05).For lower-risk patients,urgent endoscopy was independently associated with a higher likelihood of rebleeding(adjusted OR:1.73,95%CI:1.03-2.88),while it was not associated with in-hospital mortality.However,the urgent need for endoscopy was not associated with rebleeding and in-hospital mortality in high-risk patients.Conclusion:Endoscopy within 12 h did not provide any advantage in the outcomes of patients with NVUGIH,and may even lead to an increased rebleeding rate in lower-risk patients.

Chinese Trauma Surgeon Association for management guidelines of vacuum sealing drainage application in abdominal surgeries-Update and systematic review

Vacuum sealing drainage (VSD) is frequently used in abdominal surgeries. However, relevant guidelines are rare. Chin ese Trauma Surge on Associati on orga nized a committee composed of 28 experts across China in July 2017, aiming to provide an evidence-based recommendation for the application of VSD in abdominal surgeries.Eleven questions regarding the use of VSD in abdominal surgeries were addressed:(1) which type of materials should be respectively chosen for the intraperitoneal cavity, retroperitoneal cavity and superficial incisions?(2) Can VSD be preventively used for a high-risk abdominal incision w让h primary suture?(3) Can VSD be used in severely contaminated/infected abdominal surgical sites?(4) Can VSD be used for temporary abdominal cavity closure under some special conditions such as severe abdominal trauma, infection, liver transplantation and intra-abdominal volume increment in abdominal compartment syndrome?(5) Can VSD be used in abdominal organ inflammation, injury, or postoperative drainage?(6) Can VSD be used in the treatment of intestinal fistula and pancreatic fistula?(7) Can VSD be used in the treatment of intra-abdominal and extra-peritoneal abscess?(8) Can VSD be used in the treatment of abdominal wall wounds, wound cavity, and defects?(9) Does VSD in crease the risk of bleeding?(10) Does VSD increase the risk of intestinal wail injury?(11) Does VSD increase the risk of peritoneal adhesion? Focusing on these questions, evidence-based recommendations were given accordingly. VSD was strongly recommended regarding the questions 2-4. Weak recommendations were made regarding questions 1 and 5-11. Proper use of VSD in abdominal surgeries can lower the risk of infection in abdominal incisions with primary suture, treat severely contaminated/infected surgical sites and facilitate temporary abdominal cavity closure.

A convenient,highly-efficient method for preparation of hydroxyl-terminated isotactic poly(propylene) and functional di-block copolymer

Both terminated functional isotactic polypropylene （iPP） and block copolymers containing iPP segment are desirable for commercial applications. This paper provides a convenient, highly-efficient method to prepare hydroxyl-terminated isotactic polypropylene （iPP-t-OH） and functional di-block copolymer containing the iPP segment through a combination of coordination polymerization and coupling reaction. The coordination polymerization was catalyzed by TiCI4/MgCI2/AIEt3 catalyst system using ZnEt2 as chain transfer agent. Further, the Zn-terminated iPP was oxidized and subsequently hydrolyzed to provide iPP-t-OH. Soxhlet extraction and 13C NMR were used to calculate the isotacticity of iPP-t-OH. The degree of polymerization and the number of hydroxyl groups at the chain end of iPP-t-OH were measured by GPC and 1H NMR. Despite the high molecular weight and heterogeneous reaction, iPP-t-OH is effectively linked with PEG-t-NCO to produce di-block copolymers. DSC analysis of the di-block copolymer shows an obvious decrease in Tm and To, which indicated that PEG was successfully linked to the terminal end of iPP.