Objective To investigate the therapeutic effect of Yudantong decoction in mice with α-naphthyl isothiocyanate (ANIT)-induced cholestasis, as well as its targets and mechanism based on intestinal flora and intestinal barrier function. Methods A total of 24 C57BL/6 mice were randomly divided into control group, model group, Yudantong decoction group (YDTF group), and ursodeoxycholic acid (UDCA) group, with 6 mice in each group. The mice in the model group, the YDTF group, and the UDCA group were given ANIT 35 mg/kg/day by gavage on days 1, 4, 7, 10, and 13, and those in the YDTF group and the UDCA group were given Yudantong decoction or UDCA by gavage for 15 consecutive days; related samples were collected on day 16. Liver histopathology was observed, and liver function parameters were measured; immunohistochemistry was used to measure the protein expression levels of caspase-1, interleukin-1β (IL-1β), and FXR in the liver, and flow cytometry was used to measure the percentages of CD11b + , CD86 + , and CD45 + immune cells in the liver; 16S rDNA sequencing and information analysis were performed for fecal microorganisms; immunohistochemistry was used to measure the protein expression of the intestinal FXR/NLRP3 pathway, and immunofluorescence assay was used to measure the protein expression of intestinal E-cadherin and occludin. A one-way analysis of variance was used for comparison of continuous data with homogeneity of variance between multiple groups, and the least significant difference t -test was used for further comparison between two groups; the Welch test was used for comparison of data with heterogeneity of variance between multiple groups, and the Games-Howell test was used for further comparison between two groups. Results HE staining showed that the model group had partial hepatocyte fatty degeneration, massive necrosis of hepatocytes in hepatic lobules, damage of lobular structure, and massive inflammatory cell infiltration, and the YDTF group and the UDCA group had alleviation of hepatocyte fatty degeneration and hepatocyte necrosis in hepatic lobules, with a reduction in inflammatory cells. Compared with the control group, the model group had significantly higher serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transpeptidase (GGT), alkaline phosphatase (ALP), total bilirubin (TBil), direct bilirubin (DBil), and total bile acid (TBA) (all P < 0.05); compared with the model group, the YDTF group had significant reductions in the serum levels of ALT, AST, GGT, ALP, TBil, DBil, and TBA (all P < 0.05), and the UDCA group had significant reductions in the serum levels of GGT, TBil, DBil, and TBA (all P < 0.05). Compared with the control group, the model group had significant increases in the levels of caspase-1 and IL-1β and a significant reduction in the expression of FXR in the liver (all P < 0.05); compared with the model group, the YDTF group had significant reductions in the levels of caspase-1 and IL-1β in the liver and the UDCA group had a significant reduction in the level of IL-1β in the liver, and both the YDTF group and the UDCA group had a significant increase in the expression level of FXR in the liver (all P < 0.05). The model group had a significant change in the composition of intestinal flora compared with the control group ( P < 0.05); there was a significant difference in the structure of intestinal flora between the YDTF group and the model group ( P < 0.05), and there was also a significant difference in the composition of intestinal flora between the UDCA group and the control/model groups ( P < 0.05). Compared with the control group, the model group had a significant increase in the abundance of intestinal Akkermansia muciniphila and a significant reduction in the abundance of Lactobacillus johnsonii (both P < 0.05); compared with the model group, both the YDTF group and the UDCA group had a significant reduction in the abundance of intestinal Akkermansia muciniphila , and the YDTF group had a significant increase in the abundance of Lactobacillus murinus , while the UDCA group had significant increases in the abundance of Lactobacillus murinus and Bifidobacterium pseudolongum (all P < 0.05). Compared with the control group, the model group had a significant reduction in the protein expression of intestinal FXR, a significant increase in the protein expression of intestinal NLRP3, and significant reductions in the expression of intestinal E-cadherin and occludin (all P < 0.05); compared with the model group, both the YDTF group and the UDCA group had a significant increase in the protein expression of intestinal FXR, a significant reduction in the protein expression of intestinal NLRP3, and significant increases in the expression of intestinal E-cadherin and occludin (all P < 0.05). Conclusion Yudantong decoction can alleviate liver injury in mice with ANIT-induced cholestasis, possibly by improving intestinal flora and enhancing intestinal barrier function.

Objective:This study aimed to explore the feasibility and clinical value of monitoring the progression of early kidney injury in type 2 diabetic patients by assessment of the urinary C-terminal agrin fragment (uCAF) with enzymatic chemiluminescence immunoassay.Methods:A total of 251 patients with type 2 diabetes, who attended the Second Affiliated Hospital of Wenzhou Medical University from October 2018 to March 2020, were included in this retrospective analysis. One hundred and fifty-six participants undergoing health check-up at the Second Affiliated Hospital of Zhejiang University School of Medicine in February 2021 served as controls. Basic clinical information, glycosylated hemoglobin type A 1c and serum creatinine values were recorded, and urine specimens were collected for urinary creatinine, urinary α 1 microglobulin(uα 1M), urinary immunoglobulin G (uIgG), urinary albumin, urinary N-Acetyl-B-D-glycosaminidase (uNAG) and uCAF measurements. Based on the estimated glomerular filtration rate (eGFR), 251 patients were classified into G1~G5 stage groups with 116, 22, 28, 55 and 30 patients in each group. One hundred and sixty-six patients with early diabetic kidney disease (stage G1-G3) were divided into subgroups A1 (79), A2 (48) and A3 (39) according to the urinary albumin/creatinine ratio (UACR), the uα1M levels were divided into uα1M subgroup 1 (83 cases), uα1M subgroup 2 (42 cases), and uα1M subgroup 3 (41 cases), and uIgG subgroup 1 (83 cases), uIgG subgroup 2 (42 cases), and uIgG subgroup 3 (41 cases) according to uIgG levels. The Spearman method was used to analyze the correlation between uCAF levels and eGFR, UACR, uα1M and uIgG levels. Results:(1) The linear range of the uCAF detected by enzymatic chemiluminescence immunoassay was 3.97-2 000.00 ng/ml, with a detection limit of 2.28 ng/ml, intra-batch coefficients of variation of 1.15% and 1.57%, inter-batch coefficients of variation of 1.63% and 5.78%, and a biological reference interval of <95.35 μg/g Cr. (2) The uCAF level and positive rate (UACR≥30 mg/g) increased with the decrease of eGFR from G1-G3, uCAF level was negatively correlated with eGFR value ( r=-0.543, P<0.000 1), and the positive rate increased from 24.14% (28/116) to 85.71% (24/28) from G1-G3. The uCAF level and positivity rate decreased with the decrease of eGFR from G4 to G5. uCAF level was positively correlated with eGFR value ( r=0.495, P<0.001), and the positivity rate decreased from 30.91% (17/55) to 23.33% (7/30) from G4 to G5. (3) In patients with early diabetic kidney disease, uCAF levels and positivity rates increased gradually with the increase of UACR. uCAF levels were positively correlated with UACR values ( r=0.602, P<0.001), and the uCAF positivity rate reached 21.52% (17/79) in the A1 subgroup. (4) uCAF level was positively correlated with uα1M and uIgG levels in patients with early diabetic kidney disease ( r=0.757, 0.596, both P<0.001). Conclusion:Analytical performance of enzyme chemiluminescence immunoassay for the detection of CAF is satisfactory and could be used a biomarker for monitoring damage and progression of early diabetic kidney disease in patients with type 2 diabetes.

COVID-19 pandemic caused by SARS-CoV-2 infection severely threatens global health and economic development. No effective antiviral drug is currently available to treat COVID-19 and any other human coronavirus infections. We report herein that a macrolide antibiotic, carrimycin, potently inhibited the cytopathic effects (CPE) and reduced the levels of viral protein and RNA in multiple cell types infected by human coronavirus 229E, OC43, and SARS-CoV-2. Time-of-addition and pseudotype virus infection studies indicated that carrimycin inhibited one or multiple post-entry replication events of human coronavirus infection. In support of this notion, metabolic labelling studies showed that carrimycin significantly inhibited the synthesis of viral RNA. Our studies thus strongly suggest that carrimycin is an antiviral agent against a broad-spectrum of human coronaviruses and its therapeutic efficacy to COVID-19 is currently under clinical investigation.

Objective:To explore clinical manifestations, imaging features and prognosis of juvenile idiopathic arthritis (JIA) combined with lung injury, aiming to improve the understanding of the disease.Methods:Clinical data from 464 children with JIA who were hospitalized in Beijing Children′s Hospital from January 2016 to September 2019 were retrospectively analyzed.Their clinical manifestations, high resolution CT (HRCT) features, lung function and follow-up of children with lung injury were analyzed.Results:Among 464 children with JIA, 40 cases (8.62%) combined with lung injury.There were no significant differences in the age and sex between JIA children either combined with lung injury or not (all P>0.05). Among them, there were 125 cases of systemic-onset juvenile idiopathic arthritis (SoJIA) and 28 cases (22.4%) of JIA combined with lung injury, accounting for the highest proportion (70%, 28/40 cases) in JIA children combined with lung injury.Among 40 JIA children combined with lung injury, 22 cases (55.0%) had respiratory symptoms and 7 cases (17.5%) had obvious hypoxia.HRCT examination was performed in them, and the imaging findings included high-density strip or strip flocculation (75.0%, 30/40 cases), pleural thickening (45.0%, 18/40 cases), ground glass shadow (22.5%, 9/40 cases), nodular lesion (20.0%, 8/40 cases), vesicles or cystic emphysema (15.0%, 6/40 cases). Lung function was detected in 12/20 children with varying degrees of pulmonary function abnormalities, most of which were mixed ventilation dysfunction, and 2 cases still had pulmonary function abnormalities after treatment.During the follow-up for 3 months to 3.5 years, 4 cases (10%) JIA combined with lung injury died.A total of 29 children were re-examined by pulmonary CT in the follow-up visit, including 14 (48.28%) improved, 8 cases (27.58%) with no significant improvement, and 7 cases (24.14%) with repeated disease. Conclusions:JIA is a common rheumatic immune disease in children and all subtypes can be combined with lung injury, manifesting as interstitial lung disease mainly.The age and sex of JIA children combined with lung injury are not specific factors.The proportion of lung injury in SoJIA is significantly higher than that in other subtypes.SoJIA combined with macrophage activation syndrome can lead to respiratory failure, respiratory distress syndrome, and even death, which is one of the main factors leading to poor prognosis of JIA.HRCT is more sensitive to the diagnosis of lung injury.Lung function detection is a simple and easy method to evaluate and monitor lung injury.The prognosis of JIA children combined with lung injury is poor, which should be well concerned.

Mitochondrial diseases are maternally inherited heterogeneous disorders that are primarily caused by mitochondrial DNA (mtDNA) mutations. Depending on the ratio of mutant to wild-type mtDNA, known as heteroplasmy, mitochondrial defects can result in a wide spectrum of clinical manifestations. Mitochondria-targeted endonucleases provide an alternative avenue for treating mitochondrial disorders via targeted destruction of the mutant mtDNA and induction of heteroplasmic shifting. Here, we generated mitochondrial disease patient-specific induced pluripotent stem cells (MiPSCs) that harbored a high proportion of m.3243A>G mtDNA mutations and caused mitochondrial encephalomyopathy and stroke-like episodes (MELAS). We engineered mitochondrial-targeted transcription activator-like effector nucleases (mitoTALENs) and successfully eliminated the m.3243A>G mutation in MiPSCs. Off-target mutagenesis was not detected in the targeted MiPSC clones. Utilizing a dual fluorescence iPSC reporter cell line expressing a 3243G mutant mtDNA sequence in the nuclear genome, mitoTALENs displayed a significantly limited ability to target the nuclear genome compared with nuclear-localized TALENs. Moreover, genetically rescued MiPSCs displayed normal mitochondrial respiration and energy production. Moreover, neuronal progenitor cells differentiated from the rescued MiPSCs also demonstrated normal metabolic profiles. Furthermore, we successfully achieved reduction in the human m.3243A>G mtDNA mutation in porcine oocytes via injection of mitoTALEN mRNA. Our study shows the great potential for using mitoTALENs for specific targeting of mutant mtDNA both in iPSCs and mammalian oocytes, which not only provides a new avenue for studying mitochondrial biology and disease but also suggests a potential therapeutic approach for the treatment of mitochondrial disease, as well as the prevention of germline transmission of mutant mtDNA.
Animals ; DNA, Mitochondrial ; genetics ; Humans ; Induced Pluripotent Stem Cells ; cytology ; metabolism ; MELAS Syndrome ; genetics ; Male ; Mice ; Microsatellite Repeats ; genetics ; Mitochondria ; genetics ; metabolism ; Mutation ; genetics

Objective To observe the effect of Hawthorn Jiangzhi powder on blood lipids in hyperlipidemia patients.Methods Four hundreds and eighty-four patients with hyperlipidemia were selected from Department of Cardiology in Huanghuai University Affiliated Hospital from January 2011 to June 2016,and they were divided into observation group and control group by random number table,each group 242 cases.The observation group took orally Hawthorn Jiangzhi powder (including ingredients:hawthorn 6 g,salvia miltiorrhiza 18 g,black soybean 16 g,hoelen 6 g,ganoderma lucidum 9 g,kudzuvine root 6 g,Chinese yam 6 g,fructus amomum 9 g,coix seed 16 g,cassia seed 6 g) once 6-9 g powder,twice a day,once in the morning and another in the evening;the control group was given simvastatin,20 mg each day during taking dinner;the therapeutic period lasted 2 months in both groups.The differences in serum lipid and serum inflammatory factor levels were compared before and after treatment in the two groups;the changes of lymphocyte subsets of the two groups were observed and compared with the changes of the subset results of 100 normal healthy subjects aged 35-80 years old in the same period in our hospital,and the total efficiency,the situations of adverse reactions and liver and kidney functions of two groups were observed.Results In the observation group and the control group,before treatment the levels of CD4+,CD8+,CD4+/CD8+ were lower than those of healthy control group,but after treatment the levels of CD4+,CD8+ and CD4+/CD8+ were higher than those before treatment,and the changes of the observation group were more significant than those of the control group(CD4+:0.47±0.11 vs.0.40±0.10,CD8+:0.28 ± 0.10 vs.0.26 ± 0.08,CD4+/CD8+:1.67 ± 0.79 vs.1.53 ± 0.45);After treatment,the levels of hypersensitive C-reactive protein (hs-CRP),interleukin-6 (IL-6),von Willebrand factor (vWF) and homocysteine (Hcy) in the two groups were significantly lower than those before treatment,and the levels of hs-CRP,IL-6 and vWF in the observation group were obviously lower than those in the control group [hs-CRP (mg/L):5.1 ± 1.8 vs.5.8 ± 1.7,IL-6 (ng/L):2.9 ± 1.6 vs.3.7 ± 1.8,vWF:(126.8 ± 12.8)％ vs.(156.5 ± 11.3)％,all P ＜ 0.05].After treatment,Hcy in the observation group was lower than that in the control group,but there was no significant difference between the two groups (μμmol/L:5.2 ± 1.8 vs.5.4 ± 2.6,P ＞ 0.05).In the observation group after treatment at each time point,the levels of total cholesterol (TC),triglyceride (TG) and low density lipoprotein cholesterol (LDL-C) were lower than those before treatment,while the levels of high density lipoprotein cholesterol (HDL-C) and high density lipoprotein cholesterol/total cholesterol (HDL/TC) were higher than those before treatment;after treatment in the control group,the levels of TC,TG and LDL-C were decreased,and the levels of HDL-C and HDL/TC were obviously increased compared with those before treatment;The levels of TC,TG and LDL-C in the observation group after treatment for 2 months were significantly lower than those in the control group [TC (mmol/L):1.26 ± 0.57 vs.2.26 ± 0.56;TG (mmol/L):3.45 ± 0.78 vs.5.45 ± 0.75,LDL-C (mmol/L):2.40±0.65 vs.2.72±0.85;all P ＜ 0.05),and HDL/TC was obviously increased (1.19±0.15 vs.0.62±0.35,P ＜ 0.01).The total therapeutic effective rate of the observation group was significantly higher than that of the control group [90.1％ (218/242) vs.73.4％ (178/242),P ＜ 0.01].Adverse reactions and changes of liver and kidney functions during the period of treatment in the two groups were minimal.Conclusions Hawthorn Jiangzhi powder can effectively reduce the blood lipids and serum inflammation cytokines in patients with hyperlipidemia,improve blood rheological situation,reduce serum levels of inflammatory factors,inhibit the formation and development of atherosclerosis and enhance the immune function obviously in patients with high lipid abnormalities.

Objective To investigate the protective effects of rhein lysinate ( RHL) on cardiac tissue damage in-duced by paraquat in experimental mice , and to clarify its mechanism .Methods In this study mice were assigned to the following three groups: control, paraquat model, and RHL-treated groups.The model of oxidative damage mice was established by intraperitoneal injection of paraquat .RHL-treated group was given RHL ( 50 mg/kg ) by gavage for one week before performing model .The other two groups were given equal volume of distilled water .For making model , paraquat was intraperitoneally injected in the paraquat model and RHL-treated group .The content of MDA was detected by thiobarbituric acid assay .The activities of SOD and GSH-Px were detected by biphenyl three phenolic autoxidation assay and NADPH coupling method respectivly .The pathological profile of cardiac tis-sue was observed by hematoxylin and eosin ( HE) staining and reactive oxygen species was observed by DCFH-DA staining .The change of proteins related to myocardial damage detected by Western blot .Results Compared with control group, the activities of SOD and GSH-Px decreased (P<0.05) and the content of MDA increased (P<0.05) in paraquat model group .However , these changes were attenuated byr RHL treatmen ( P<0.05 ) .The pathologi-cal examination indicated the structure of cardiac tissue was damaged and reactive oxygen species of cardiac tissue was increased after paraquat was given , however , these changes were attenuated after RHL treatmen .It was shown in western blot analysis that compared with control group , the expression of SIRT1 decreased, the acetylation of P53 and the expression of P 53 and P66 increased in paraquat-treated group .These changes were attenuated by RHL treatmen ( P<0.05 ) .Conclusions RHL may attenuate paraquat-induced cardiac injury in mice .

Objective To systematically evaluate the safety of laparoscopic distal pancreatectomy (LDP) compared with open distal pancreatectomy (ODP).Methods Databases including Cochrane library,MEDLINE,EMbase,Google Scholar and Chinese National Knowledge Infrastructure were searched to enroll randomized clinical trials (RCT),controlled clinical trials (CCT) or retrospective case-control studies to compare LDP with ODP.All articles received quality assessment according to the inclusion and exclusion criteria,then the selected indices were analyzed using the Review Manager Version 5.0 software (The Cochrane Collaboration,Oxford,United Kingdom).Results 21 manuscripts with a total of 2 797 patients were enrolled.1 150 patients underwent LDP and the remaining 1 647 patients underwent ODP.In 20 studies (n =2 597),the total postoperative complication rates were 33.90％ for the LDP group versus 46.80％ for the ODP group [RR =0.76,95％ CI(0.69 ～ 0.84),P ＜ 0.01].In 8 studies (n =1 869) there was no significant difference [RR =0.51,95％ CI(0.21 ～ 1.24),P ＞0.05] in the perioperative mortality between LDP (4/703) and ODP (18/1 166).In 20 studies (n =2 757) there was no significant difference [RR =0.89,95％ CI(0.75 ～ 1.06),P ＞ 0.05] in the pancreatic fistula rate between LDP (168/1 132) and ODP (281/1 625).In 11 studies (n =1 840) the wound infection rate of LDP (3.24％) was significantly lower than ODP (10.85％) [RR =0.34,95％ CI(0.23 ～ 0.52),P ＜ 0.01].No significance was found between the two groups in the rates of pulmonary complications,peritoneal infection,urinary tract infection,postoperative bleeding,pseudocyst formation,intestinal obstruction and ascites formation between LDP and ODP.Conclusions When compared with the traditional open procedure,LDP has the advantages of significantlylower rates of postoperative complication and wound infection.There were no significant differences in postoperative mortality,and pancreatic fistula rate between LDP and ODP.This meta-analysis suggests that LDP is a safe and feasible operative method.

Objective: To investigate the effect of gross total resection on the local control and survival of patients with stage IV neuroblastoma (NB) and analyze the extent of surgical resection of primary tumors that affects patient survival. Methods: A total of 96 patients with stage Ⅳ NB who were admitted to the Sun Yat-Sen University Cancer Center between January 2000 and December 2011 were analyzed. The patients were treated with combined-modality therapy, including chemotherapy, surgery, and/or radiotherapy. The patients were divided according to the extent of surgical resection of primary tumor into the following groups: group A, biopsy or tumor removal of less than 50% of the primary lesion; group B, incomplete resection of more than 50% but less than 90% of the lesion; group C, removal of more than 90% of the lesion; and group D, complete resection with or without macroscopic residual tumors. The survival rates of each group were analyzed. Results: The median age of the 96 patients was 4.4 years, ranging from 1.2-18.8 years. The overall 3-year progression-free survival (PFS) and overall survival (OS) of the total patients were 32.8% and 36.7%, respectively. A total of 24 cases were assigned in group A, 10 in group B, 23 in group C, and 39 in group D. Subgroup analysis revealed that the 3-year PFS rate was 17.5% for group A, 20.0% for group B, 45.1% for group C, and 40.5% for group D. The PFS rates were not statistically significant-ly different between groups A and B (P=0.352) and between groups C and D (P = 0.792). However, the OS was higher in groups C and D than that in groups A and B. The 3-year PFS rates were 42.2% and 17.8% for groups C and D (P<0.001), respectively. Conclu-sion: Resection extension of more than 90% of the primary tumor combined with chemotherapy and (or) radiation therapy can improve the survival of patients with stage Ⅳ NB. However, this treatment modality does not affect the treatment outcomes for minimal gross tu-mor residuals.

OBJECTIVETo evaluate the long-term survival of children and adolescents with lymphoblastic lymphoma (LBL) treated by a modified NHL-BFM-90 protocol.

METHODSFrom March 1998 to November 2010, 107 untreated patients with LBL (age <18 years) were enrolled and stratified into three groups (R1, R2 and R3), according to the stage of disease and response to induction chemotherapy. All patients received different intensive chemotherapy regimens based on a modified NHL-BFM-90 protocol. Total treatment duration was 2 years.

RESULTSOf the 107 patients, 79 were boys and 28 were girls, with a median age of 10 years (range 2.5-18 years). Six patients (5.6%) were stage I/II, 101 (94.4%) stage III/IV. The R1, R2 and R3 groups accounted for 5.6%, 71.0% and 23.4%, respectively. 75.7% of the patients had T-LBL, and 24.3% was B-LBL. At a median follow-up duration of 60 months (range 1-186 months), 24 patients died. The 5-year event-free survival (EFS) and overall survival (OS) were 75.5% and 77.8 % for all patients, 100.0% and 100.0% for group R1, 84.5% and 87.5 % for R2, 44.0% and 44.0% for R3, 72% and 73.5% for T-LBL, 86.4% and 88.5% for B-LBL, respectively. Myleosuppression was the major toxicity and need aggressive management.

CONCLUSIONThe modified NHL-BFM-90 protocol is an effective therapy for children and adolescents with LBL in low and intermediate risk. T-LBL had the similar outcomes as B-LBL did. The patients in high-risk group had a poor survival and new protocols are needed.

Adolescent ; Antineoplastic Combined Chemotherapy Protocols ; Asparaginase ; Child ; Child, Preschool ; Daunorubicin ; Disease-Free Survival ; Female ; Humans ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; Prednisone ; Treatment Outcome ; Vincristine