Effect of Lactobacillus plantarum WCFS1 on pancreatic and ileal injury in mice with acute necrotizing pancreatitis / 中华胰腺病杂志

Objective:To explore the effect of probiotics Lactiplantibacillus plantarum(LP) WCFS1 by gavage on acute necrotizing pancreatitis (ANP) and associated ileum injury in mice. Methods:Twenty-four healthy male mice were gavaged with broad-spectrum antibiotics for 3 weeks to establish microbiota-depleted mice, and then randomly divided into control group (CON), ANP model group (ANP), LP gavage group (LP) and LP gavage and ANP induced group (LP+ ANP) , with 6 mice in each group. Mice in LP and LP+ ANP group were treated by gavage of LP (1×10 9 CFU/ml, 0.2 ml/day per mouse) for 1 week, while CON and ANP were gavaged with sterile phosphate buffered saline for 1 week instead. The ANP model was induced by intraperitoneal injection with caerulein (100 μg/kg) for 10 times with 1-hour interval between two injections and the 10th injection with lipopolysaccharide(LPS) 5 mg/kg intraperitoneally, and the mice were sacrificed 2 h later. Levels of LP in stool and ileal mucosa were detected by real-time PCR; the pancreas and ileum were collected for pathological examination to observe the extent of tissue inflammation and to score the pathology. Serum amylase activities were determined by enzymatic kinetic chemistry; serum inflammators levels and intestinal permeability were detected by ELISA; levels of inflammators in pancreatic and ileal tissues were detected by real-time PCR; ileal tight-junction proteins (occludin, claudin-1 and ZO-1) were measured by immunofluorescence staining. Results:LP levels in the stool and ileal mucosa of mice were significantly increased after LP gavage, and the differences were statistically significant (913.30±39.12 vs 2.39±1.39, 23.11±0.50 vs 1.38±0.28, all P value <0.05). The pathological scores of pancreatic tissue of CON, LP, ANP and LP+ ANP group were (0.26±0.41), (0.17±0.26), (8.55±0.46) and (6.30±0.45); the serum amylase activities were (219.70±19.73), (217.60±11.30), (2896.24±98.32) and (1837.13±131.60)U/L, IL-1β were (0.87±0.28), (1.4±0.85), (67.41±6.45) and (36.33±5.65)pg/ml, IL-6 were (0.74±0.27), (0.16±0.16), (280586.12±39163.92) and (107912.75±31283.47)pg/ml, IL-10 were (35.52±5.27), (50.99±15.34), (2008.45±184.83) and (3070.35±403.71)pg/ml; the expression level of pancreatic IL-1β mRNA was 1.42±0.39, 0.95±25, 20.53±0.50 and 10.69±1.01, IL-6 mRNA was 1.31±0.44, 0.93±0.023, 21.97±1.71 and 11.54±1.75, IL-10 mRNA was 0.93±0.14, 0.75±0.15, 0.99±0.21 and 1.76±0.19; there was no significant difference between LP and CON group, and pancreatic pathological scores, serum amylase、IL-1β and IL-6 levels, and the expression level of pancreatic IL-1β and IL-6 mRNA were significantly decreased in LP+ ANP group compared with those in ANP group, while serum IL-10 levels and the expression level of pancreatic IL-10 mRNA were significantly increased compared with ANP group, and all the differences were statistically significant (all P values <0.05). The pathological scores of ileal tissue of CON, LP, ANP and LP+ ANP group were 0, 0, (3.17±0.41) and (1.67±0.52); the levels of serum DAO of CON, LP, ANP and LP+ ANP group were (0.03±0.03), (0.02±0.02), (0.50±0.05) and (0.49±0.06)ng/ml; LPS levels were (2.75±0.35), (3.74±0.28), (7.19±0.92) and (5.88±0.38)ng/ml; the expression level of ileal IL-1β mRNA was 1.21±0.20, 1.17±0.09, 1.81±0.25 and 1.63±0.21; IL-6 mRNA was 1.01±0.29, 2.83±0.42, 54.45±8.50 and 16.87±4.42; IL-10 mRNA was 1.12±0.41, 6.09±2.51, 11.65±1.47 and 29.86±2.93. There was no significant difference between LP and CON group, except that the ileal IL-10 mRNA expression was significantly higher than that of CON group. Ileal pathological scores, serum LPS levels and the expression level of ileal IL-6 mRNA were significantly lower in LP+ ANP group than those in ANP group, while the expression level of ileal IL-10 mRNA was significantly higher than that of ANP group; the expression of ileal tight junction proteins (ocludin, claudin-1, ZO-1) was significantly higher than those in ANP group, and all the differences were statistically significant (all P values <0.05). Conclusions:LP WCFS1 gavage could ameliorate the injury of pancreatic and ileal barrier in caerulein-induced ANP mice.

Establishment of mouse pancreatic ductal organoids and exploration of its physiological functions / 中华胰腺病杂志

Objective:To establish culture system for mouse pancreatic ductal organoids and investigate the morphology and physiological functions of the organoids.Methods:Pancreatic tissues were taken from C57BL/6 mice (6-8 weeks) and digested by collagenase Ⅳ. The pancreatic ducts were separated and collected and then the pancreatic organoids were cultured in the complete medium after Matrix gel embedding. Morphological evaluation of the organoids was performed after hematoxylin-eosin staining. The expression and localization of markers for organoids were identified by Western Blot and immunofluorescence staining; and the expression and localization of ion channels and antimicrobial peptides of the organoids were detected by agarose gel electrophoresis and immunofluorescence staining.Results:Mouse pancreatic organoids were successfully established, which could be stably passaged for 10 generations. The organoids grew spherically and formed a duct-like structure. The internal cavity corresponded to the lumen of pancreatic duct tissue. The pancreatic organoids stably expressed stem progenitor cell marker gene SOX9 and ductal epithelial cell-specific gene KRT19, which were both localized in the epithelium. The organoids did not express amylase. The organoids maintained stable expression of epithelial ion channels Clcn1, Kcnma1, CFTR, Slc12a5, Slc26a3, Slc26a6 and Scnn1a, low expression of Ano1 and no expression of Clcn3, Kcna1, Kcna2, Kcnd3, Kcnh1, Atp12a, Slc4a4, Slc9a1, Slc12a2 and Slc26a11; and CFTR highly expressed in epithelial cells. The organoids maintained high expression of antimicrobial peptides Reg3a, CRAMP and glycoprotein 2, low expression of Defb1, Defb2, and Defb3 and no expression of Defa1 and Defa4; and both CRAMP and Reg3a were expressed in the epithelial cells and secreted into the lumen of the organoids.Conclusions:Mouse pancreatic organoids are successfully established, which can be stably passaged. The organoids maintain the characteristics of ductal epithelial cells and can be used as an in vitro model to study the physiology of pancreatic ducts.

Immunometabolism: a new dimension in immunotherapy resistance / 医学前沿

Immune checkpoint inhibitors (ICIs) have demonstrated unparalleled clinical responses and revolutionized the paradigm of tumor treatment, while substantial patients remain unresponsive or develop resistance to ICIs as a single agent, which is traceable to cellular metabolic dysfunction. Although dysregulated metabolism has long been adjudged as a hallmark of tumor, it is now increasingly accepted that metabolic reprogramming is not exclusive to tumor cells but is also characteristic of immunocytes. Correspondingly, people used to pay more attention to the effect of tumor cell metabolism on immunocytes, but in practice immunocytes interact intimately with their own metabolic function in a way that has never been realized before during their activation and differentiation, which opens up a whole new frontier called immunometabolism. The metabolic intervention for tumor-infiltrating immunocytes could offer fresh opportunities to break the resistance and ameliorate existing ICI immunotherapy, whose crux might be to ascertain synergistic combinations of metabolic intervention with ICIs to reap synergic benefits and facilitate an adjusted anti-tumor immune response. Herein, we elaborate potential mechanisms underlying immunotherapy resistance from a novel dimension of metabolic reprogramming in diverse tumor-infiltrating immunocytes, and related metabolic intervention in the hope of offering a reference for targeting metabolic vulnerabilities to circumvent immunotherapeutic resistance.

One-step synthesis of site-specific antibody-drug conjugates by reprograming IgG glycoengineering with LacNAc-based substrates

Glycosite-specific antibody‒drug conjugatess (gsADCs), harnessing Asn297 N-glycan of IgG Fc as the conjugation site for drug payloads, usually require multi-step glycoengineering with two or more enzymes, which limits the substrate diversification and complicates the preparation process. Herein, we report a series of novel disaccharide-based substrates, which reprogram the IgG glycoengineering to one-step synthesis of gsADCs, catalyzed by an endo-N-acetylglucosaminidase (ENGase) of Endo-S2. IgG glycoengineering via ENGases usually has two steps: deglycosylation by wild-type (WT) ENGases and transglycosylation by mutated ENGases. But in the current method, we have found that disaccharide LacNAc oxazoline can be efficiently assembled onto IgG by WT Endo-S2 without hydrolysis of the product, which enables the one-step glycoengineering directly from native antibodies. Further studies on substrate specificity revealed that this approach has excellent tolerance on various modification of 6-Gal motif of LacNAc. Within 1 h, one-step synthesis of gsADC was achieved using the LacNAc-toxin substrates including structures free of bioorthogonal groups. These gsADCs demonstrated good homogeneity, buffer stability, in vitro and in vivo anti-tumor activity. This work presents a novel strategy using LacNAc-based substrates to reprogram the multi-step IgG glycoengineering to a one-step manner for highly efficient synthesis of gsADCs.

Research Progress of Immunotherapy for Non-small Cell Lung Cancer with Drive Gene Mutation / 中国肺癌杂志

Lung cancer is the most lethal malignancy around the world and non-small cell lung cancer (NSCLC) accounts for 80% of all cases. Most of the NSCLC patients has "driver gene mutations" and targeted therapy achieved a relatively good efficacy, but some patients progressed or relapsed after treatment. Previous studies demonstrated that immune checkpoint inhibitor could improve the prognosis of advanced-stage NSCLC and prolong the survival time. However, the efficacy of immune therapy varies in NSCLC patients with different immune and molecular features. The efficacy of immune therapy was controversial in NSCLC patients with driver gene mutation. The present review will summarize the immune characteristics of NSCLC patients with driver mutation and the directions of immunotherapy for patients with driver mutation.
.

C-Reactive Protein, Interleukin-6 and Procalcitonin in Early Assessment of Severity of Acute Pancreatitis / 胃肠病学

Background: Acute pancreatitis (AP) is a common acute abdominal disease in clinical practice. Severe AP (SAP) usually progresses rapidly, leading to systemic inflammatory response syndrome, multiple organ failure, and even death. Early identification of severe cases with timely and effective treatment can improve patient's prognosis and reduce mortality. Aims: To explore the value of single and combined detection of C-reactive protein (CRP), interleukin-6 (IL-6) and procalcitonin (PCT) in the early assessment of severity of AP. Methods: A total of 472 AP patients hospitalized in Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine from January 2018 to December 2020 were enrolled retrospectively. The laboratory data within 48 h of admission were extracted, and the patients were allocated into mild AP (MAP) group and SAP group (including moderately severe and severe AP) according to the 2012 revised Atlanta classification. The differences of CRP, IL-6 and PCT between the two groups were statistically analyzed, and the value of single and combined detection of the three indicators for predicting the disease severity was determined by ROC curve analysis. Results: The levels of CRP, IL-6 and PCT in SAP group were significantly higher than those in MAP group (all P< 0.05). ROC curve analysis revealed that the area under the curve (AUC) of single, two and three indicators combined for predicting SAP was all higher than 0.7, which represented a moderately diagnostic accuracy. Three indicators combined demonstrated the highest diagnostic accuracy (AUC=0.826); with an optimal cut-off value, the sensitivity and specificity were 71.4% and 79.3%, respectively. Conclusions: CRP, IL-6 and PCT are meaningful in early assessment of severity of AP. The overall diagnostic value of combined detection of three indicators is superior to single and two indicators combination in predicting SAP.

Update on the treatment of childhood-onset systemic lupus erythematosus / 中华实用儿科临床杂志

Systemic lupus erythematosus (SLE) is a serious multisystem autoimmune disease with different clinical manifestations.Childhood-onset SLE (cSLE) is similar to adult-onset SLE, while its morbidity and mortality are higher than adults, and it is prone to damage important organs.Therefore, early diagnosis and treatment are very important.With the in-depth exploration of the pathogenesis and the development of cell and molecular biology, the progress of drug therapy for SLE has been promoted.Immunosuppression still remains the cornerstone of treatment, and glucocorticoids still plays a leading role.Biologics bring the gospel to SLE patients, and non-specific immunotherapy gains treatment time for refractory and severe SLE patients.Treatment options are led by the level of disease severity.It is of great significance to understand the treatment progress of cSLE and combine theory with practice together to control the disease activity and improve the prognosis.This article reviews recent advances regarding the update on the treatment of cSLE in recent years.

Recent advances in the diagnosis and management of Takayasu arteritis complicated with ocular ischemic syndrome / 中华眼底病杂志

Takayasu arteritis (TA) is a rare, chronic non-specific vasculitis that can lead to ocular hypoperfusion. As a result, ocular ischemic syndrome (OIS), which prominently manifests as Takayasu retinopathy, may develop subsequently. Ocular manifestations mainly consist of progressive painless vision loss and amaurosis fugax in TA patients complicated with OIS. However, due to the lack of specific clinical characteristics, it is of great significance to improve the timely diagnosis aided by multimodal imaging, especially fundus fluorescein angiography. Early diagnosis of OIS is essential for reversal of ocular ischemia and better prognosis of TA patients. Management of OIS patients associated with Takayasu arteritis requires a combination of systemic and ophthalmic treatment. Therefore, the optimal individualized regiment should be determined by disease activity and progression, which are addressed by multi-disciplinary team assessment. Ophthalmologists should further understand the clinical features and the importance of regular ophthalmologic examinations during the disease course, thus to improve the overall survival and visual outcomes.

Research of TLD calibrated in different standard X-ray RQR radiation field in terms of eye lens dose Hp(3) / 中华放射医学与防护杂志

Objective@#To compare the calibration result of standard X-ray RQR radiation field between SSDL (NIRP) and CEA LIST LNHB(France), and to explore the feasibility of calibrating Hp(3) in standard X-ray RQR radiation field of SSDL(NIRP).@*Methods@#Using a column model with a diameter and high of 20 cm, TLD was calibrated in SSDL (NIRP) and CEA LIST LNHB (France) to measure the personal dose equivalent eye lens dose Hp(3), X-ray RQR radiation field included RQR4(60 kV), RQR7(90 kV), RQR9(120 kV), with energy response, angle response and linear response.@*Results@#In terms of energy response, the calibration results of TLD in both SSDL (NIRP) and CEA LIST LNHB (France) were in good agreement. The difference between exposure value and response value was less than 10%. In terms of angle response, the calibration result of TLD in CEA LIST LNHB (France) was better in SSDL(NIRP). The difference between exposure value and response value in CEA LIST LNHB (France) was less than 6%, while the difference between exposure value and response value in SSDL(NIRP) was more than 10% at angle of 20°. In terms of linear response, both calibration result of SSDL (NIRP) and CEA LIST LNHB (France) were in good agreement.@*Conclusions@#The standard X-ray RQR field in SSDL (NIRP) can be used for the calibration of Hp(3).

Research of TLD calibrated in different standard X-ray RQR radiation field in terms of eye lens dose Hp (3) / 中华放射医学与防护杂志

Objective To compare the calibration result of standard X-ray RQR radiation field between SSDL (NIRP) and CEA LIST LNHB (France),and to explore the feasibility of calibrating Hp (3) in standard X-ray RQR radiation field of SSDL (NIRP).Methods Using a column model with a diameter and high of 20 cm,TLD was calibrated in SSDL (NIRP) and CEA LIST LNHB (France) to measure the personal dose equivalent eye lens dose Hp (3),X-ray RQR radiation field included RQR4 (60 kV),RQR7 (90 kV),RQR9 (120 kV),with energy response,angle response and linear response.Results In terms of energy response,the calibration results of TLD in both SSDL (NIRP) and CEA LIST LNHB (France) were in good agreement.The difference between exposure value and response value was less than 10％.In terms of angle response,the calibration result of TLD in CEA LIST LNHB (France) was better in SSDL (NIRP).The difference between exposure value and response value in CEA LIST LNHB (France) was less than 6％,while the difference between exposure value and response value in SSDI (NIRP) was more than 10％ at angle of 20°.In terms of linear response,both calibration result of SSDL (NIRP) and CEA LIST LNHB (France) were in good agreement.Conclusions The standard X-ray RQR field in SSDL (NIRP) can be used for the calibration of Hp (3).

Effect of exosomes from human adipose-derived stem cells on wound healing and migration potential of HaCaT cells: a preliminary experimental study / 中华皮肤科杂志

Objective To preliminarily evaluate the effect of exosomes from human adipose-derived stem cells (hADSCs) on the migration of epidermal cells.Methods hADSCs were isolated from adipose tissues obtained from a healthy woman after liposuction,and subjected to culture.Exosomes were collected from the culture supernatant of hADSCs by using a modified method combining ultracentrifugatiou with ultrafiltration,and identified and isolated by transmission electron microscopy (TEM),dynamic light scattering (DLS) and Western blot analysis.Some cultured HaCaT cells were divided into several groups to be treated with hydrogen peroxide (H2O2) at different concentrations of 0,50,100,200,250,300,400,500,600,800 μmol/L for 1 hour,and cell counting kit-8 (CCK8) assay was performed to evaluate the effect of H2O2 on the survival rate of HaCaT cells.Some HaCaT cells were classified into 2 groups to be pretreated with phosphate-buffered saline (PBS) (normal group) or 100 μmol/L H2O2 (injury group) for 0.5 hour;then,HaCaT cells in the 2 groups were separately divided into treatment group and control group to be cocultured with exosomes or not.Confocal fluorescence microscopy was conducted to confirm the uptake of PKH26-labelled exosomes by HaCaT cells,scratch assay to estimate the wound healing potential,and Transwell assay to evaluate the migratory activity of HaCaT cells.Statistical analysis was carried out by using one-way analysis of variance for comparison among groups,least significant difference (LSD)-t test for multiple comparisons,and Spearman correlation analysis for analyzing correlations.Results TEM showed that the exosomes isolated from hADSCs were saucer-like nanovesicles with diameters of 60-80 nm.DLS revealed that the purity of the isolated exosomes was 65.88％,and they were stained positively for CD63,Alix and TSG101,which coincided with the basic characteristics of exosomes.CCK8 assay showed that survival rates of HaCaT cells gradually decreased along with the increase of H2O2 concentrations after 1-hour treatment,and were negatively correlated with the concentration of H2O2 (r =-0.91,P ＜ 0.01).Confocal fluorescence microscopy showed that the isolated exosomes could be endocytosed into impaired HaCaT cells.Scratch assay showed that the gap-filling rates at 12 hours were 40.26％ ± 0.64％,69.57％ ± 0.69％,32.28％ ± 0.31％ and 69.62％ ± 1.68％ in the normal control group,normal treatment group,injury control group and injury treatment group respectively,and the injury treatment group showed a significantly increased gap-filling rate compared with the injury control group (t =37.33,P ＜ 0.01).Transwell assay showed that the number of migratory cells per × 10 field was 20.85 ± 4.84,44.8 ± 5.24,14.95 ± 2.58 and 40.05 ± 7.66 in the normal control group,normal treatment group,injury control group and injury treatment group respectively,and was significantly larger in the injury treatment group than in the injury control group (t =25.10,P ＜ 0.01).Conclusion Exosomes isolated from hADSCs can improve the migration of HaCaT cells after oxidative injury.

Progress of diagnosis and treatment for cystic kidney disease in children / 中华实用儿科临床杂志

Cystic kidney disease is the main disease of cystic kidney change in children. It may be caused by non_genetic fetal malformations or genetic diseases,or may be acquired rarely. Most renal cysts are usually isolated oraspart of a syndrome. However,fatal renal cystic diseases can develop from these space occupying lesions. Although renalcystic diseases are similar in presentation,they possess distinct features and variable prognosis later in life. In order to correctly diagnose this kind of disease in the early stage,it claim to accurately grasp its pathogenesis,pathology,clini_cal characteristics and radiological findings. A comprehensive analysis of common cystic kidney disease in children is carried out to help clinicians to aid in early distinction and appropriate treatment.

Effect of exosomes from human adipose-derived stem cells on wound healing and migration potential of HaCaT cells: a preliminary experimental study / 中华皮肤科杂志

Objective@#To preliminarily evaluate the effect of exosomes from human adipose-derived stem cells (hADSCs) on the migration of epidermal cells.@*Methods@#hADSCs were isolated from adipose tissues obtained from a healthy woman after liposuction, and subjected to culture. Exosomes were collected from the culture supernatant of hADSCs by using a modified method combining ultracentrifugation with ultrafiltration, and identified and isolated by transmission electron microscopy (TEM) , dynamic light scattering (DLS) and Western blot analysis. Some cultured HaCaT cells were divided into several groups to be treated with hydrogen peroxide (H2O2) at different concentrations of 0, 50, 100, 200, 250, 300, 400, 500, 600, 800 μmol/L for 1 hour, and cell counting kit-8 (CCK8) assay was performed to evaluate the effect of H2O2 on the survival rate of HaCaT cells. Some HaCaT cells were classified into 2 groups to be pretreated with phosphate-buffered saline (PBS) (normal group) or 100 μmol/L H2O2 (injury group) for 0.5 hour; then, HaCaT cells in the 2 groups were separately divided into treatment group and control group to be co-cultured with exosomes or not. Confocal fluorescence microscopy was conducted to confirm the uptake of PKH26-labelled exosomes by HaCaT cells, scratch assay to estimate the wound healing potential, and Transwell assay to evaluate the migratory activity of HaCaT cells. Statistical analysis was carried out by using one-way analysis of variance for comparison among groups, least significant difference (LSD) -t test for multiple comparisons, and Spearman correlation analysis for analyzing correlations.@*Results@#TEM showed that the exosomes isolated from hADSCs were saucer-like nanovesicles with diameters of 60-80 nm. DLS revealed that the purity of the isolated exosomes was 65.88%, and they were stained positively for CD63, Alix and TSG101, which coincided with the basic characteristics of exosomes. CCK8 assay showed that survival rates of HaCaT cells gradually decreased along with the increase of H2O2 concentrations after 1-hour treatment, and were negatively correlated with the concentration of H2O2 (r= -0.91, P < 0.01) . Confocal fluorescence microscopy showed that the isolated exosomes could be endocytosed into impaired HaCaT cells. Scratch assay showed that the gap-filling rates at 12 hours were 40.26% ± 0.64%, 69.57% ± 0.69%, 32.28% ± 0.31% and 69.62% ± 1.68% in the normal control group, normal treatment group, injury control group and injury treatment group respectively, and the injury treatment group showed a significantly increased gap-filling rate compared with the injury control group (t = 37.33, P < 0.01) . Transwell assay showed that the number of migratory cells per × 10 field was 20.85 ± 4.84, 44.8 ± 5.24, 14.95 ± 2.58 and 40.05 ± 7.66 in the normal control group, normal treatment group, injury control group and injury treatment group respectively, and was significantly larger in the injury treatment group than in the injury control group (t = 25.10, P < 0.01) .@*Conclusion@#Exosomes isolated from hADSCs can improve the migration of HaCaT cells after oxidative injury.

The effects of hypertriglyceridemia on autophagy of acinar cells in acute necrotic pancreatitis rats / 中华胰腺病杂志

Objectives To observe the changes of autophagy of acinar cells in hypertriglyceridemia (HTG) associated acute necrotic pancreatitis(ANP) rats,and investigate the effects of HTG on autophagy and AP. Methods Forty SD rats were randomly divided into 4 groups using random number method, including control group,ANP group,hyperlipemia(HL) group,hyperlipidemia associated acute necrotizing pancreatitis (HANP) group. Control group and ANP group were fed with normal diet for 2 weeks,while the rats in HL and HANP group were fed with a high-fat diet for 2 weeks,and after the blood collection ANP and HANP rats were induced by 3.5% sodium taurocholate retrograde injection into the pancreaticobiliary duct to establish the ANP model. Rats in control group and HL group were intraperitoneally injected with equal volume of normal saline solution. ALL rats were sacrificed at 48 h after the model establishment. Serum triglyceride (TG), total cholesterol (TC) and serum amylase levels were measured; pancreatic tissue was histologically examined and scored; Transmission electron microscope were used to observe the intracellular ultrastructure changes and calculate the number of autophagosome and autophagic lysosome;the expressions of autophagy related proteins such as LC3,LC3Ⅱ, LC3Ⅰ, p62 and Lamp-2 were determined by Western-blot. Results After 2-week high fat diet, serum levels of TG and TC in HANP and HL groups were obviously higher than those in the control and ANP group;Serum amylase levels in HANP group and ANP group were higher than the control and HL group,and the differences were statistically significant (all P<0.05). Transmission electron microscope showed that the number of autophagosomes in ANP group was significantly higher than those in other three groups and the difference was statistically different(all P<0.05);and the autophagy activity in HANP group was impaired compared with ANP group. Compared with the control group, in ANP group the protein expressions of LC3 Ⅱ/Ⅰ ratio and p62 in pancreatic tissue were up-regulated (2.11 ± 0.03 vs 1.70 ± 0.08, 9 420 ± 624 vs 4 973 ± 577),but LAMP-2 was down-regulated(20 533 ± 578 vs 23 231 ± 1 155). Compared with ANP group,the proteins expression levels of LC3Ⅱ/Ⅰand p62 in HANP group was obviously decreased (20 533 ± 578 vs 23 231 ± 1 155), and the difference was statistically significant (all P<0.05). Conclusions Hypertriglyceridemia can worsen the pathology injury of AP,and the mechanism may be related to the decreased autophagy related proteins LC3Ⅱ/Ⅰ and p62 and decreased number of autophagosome.

Anti-inflammatory effect of interleukin-35 in mice with colitis and its mechanism / 浙江大学学报·医学版

OBJECTIVE@#To investigate the anti-inflammatory effect and mechanisms of interleukin-35 (IL-35) in inflammatory bowel disease.@*METHODS@#BALB/c mice were divided into three groups with 10 mice in each group:control group, model group (oral administration of 4% glucan sodium sulfate for 7 d) and IL-35-treated group (oral administration of 4% glucan sodium sulfate for 7 d, intraperitoneal injection of 2 μg IL-35 at d2-5). Disease activity index (DAI) was scored every day. After 7 d, the mice were sacrificed, and the serum and intestinal tissue samples were collected. The gross morphology of the colon was observed; HE staining was used to observe the pathological changes of colon tissue; flow cytometry was employed to detect the change of macrophage polarization ratio in colon tissue; the mRNA expression levels of cytokines IL-6, TNF-α, IFN-γ, IL-10 and SHIP1 in colon tissue were determined by real-time quantitative RT-PCR; the expression and distribution of SHIP1 in colon tissue was measured by immunohistochemistry; Western blotting was adopted to detect the expression level of SHIP1 protein in colonic intestinal tissues of each group.@*RESULTS@#The DAI scores of the mice in the model group were higher than those in the control group, while the DAI scores in the IL-35-treated group were lower than those in the model group (all 0.05). Compared with the model group, microscopic inflammatory infiltration score was decreased and microscopic crypt destruction and score was significantly lower in IL-35-treated group (all <0.05). The relative expression of proinflammatory cytokines IL-6, TNF-α and IFN-γ in the colon tissue of IL-35-treated group was decreased compared with the model group, while the relative expression of IL-10 mRNA was higher than that of the model group (all <0.05). Compared with the control group, the proportion of M1 macrophages in the model group increased (<0.05), while the proportion of M1 macrophages in the IL-35-treated group was lower than that in the model group (<0.05). The relative expression of SHIP1 mRNA and protein in the colon tissue of IL-35-treated group was higher than that in the model group (all <0.05).@*CONCLUSIONS@#IL-35 can inhibit the polarization of M1 macrophages and regulate inflammatory cytokines to promote anti-inflammatory effect on mice with colitis.

Effect of deoxycholic acid intervention on growth of ileum organoids derived from C57BL/6 mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish a culture system for mouse intestinal organoids and investigate the effect of deoxycholic acid (DCA) on organoids growth.</p><p><b>METHODS</b>The terminal ileum was collected from 8-month-old C57BL<6 mice. The tissue blocks were treated with EDTA and the crypts were collected and embedded in Matrigel Matrix. Orgnoids growth and buddings were observed in the control group, anhydrous alcohol group, short-term (2 days) 100 µmol<L DCA treatment group, and long-term (10 days) 10 µmol<L DCA treatment group; the orgnoids were further cultured for 10 days after removal of DCA from the medium and observed for orgnoids growth and buddings.</p><p><b>RESULTS</b>Short-term treatment with high-concentration DCA resulted in significantly reduced enterosphere formation, enteroids formation, progression from enterospheres to enteroids and number of crypt buds per enteroid (P<0.05), which remained unchanged even after removal of DCA for a short time (P<0.05); long after DCA removal, the enteroids formation rate and number of the crypt buds still remained lower than those in normal organoids (P<0.05). Short-term treatment with low-concentration DCA only resulted in reduced enteroids formation rate and number of crypt buds (P<0.05), and prolonged treatment caused reduced enterospheres formation rate, enteroids formation rate and number of crypt buds (P<0.05). After DCA removal, enteroids formation rate and the number of crypt buds still remained lower than those in the normal group (P<0.05).</p><p><b>CONCLUSION</b>We successfully established an organoids culture system. The presence of DCA in the culture system affects the growth of the organoids, which can partly recover following a prolonged period after the removal of DCA.</p>

High-throughput sequencing for analysis of structural change of intestinal microbiota in patients with colorectal adenoma / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the taxonomic richness and diversity of gut microbiota in patients with colorectal adenoma and elucidate the role of gut microorganisms in precancerous lesions in the colon and rectum.</p><p><b>METHOD</b>Adenomatous tissues from 31 patients with colorectal adenoma and normal intestinal mucosal tissues from 20 healthy control subjects were collected through colonoscopy. The total bacterial genomic DNA was extracted, and the V-Vhypervariable region in bacterial 16S rRNA gene was amplified using polymerase chain reaction and sequenced on an Illumina MiSeq platform.</p><p><b>RESULTS</b>Patients with colorectal adenomas had a higher alpha diversity and richness indices compared to the healthy controls (P<0.01). The mucosal microbiota in colorectal adenoma tissue showed a distinctive structural difference from that in normal intestinal mucosal tissues. At the phylum level, a large decrease in Firmicutes with concomitant relative expansion of Proteobacteria was observed in patients with colorectal adenomas, resulting in a significant decrease in the Firmicutes/Bacteroidetes ratio (P<0.01). At the genus level, Lactococcus and Pseudomonas were enriched whereas Enterococcus, Bacillus, and Solibacillus were reduced obviously in the preneoplastic tissues (P<0.01). We also found a similar gut microbiome composition between low-grade and high-grade intraepithelial neoplasia; the ratio of Escherichia-Shigella tended to increase in high-grade intraepithelial neoplasia, but this change was not statistically significant (P%0.28).</p><p><b>CONCLUSION</b>Significant changes in the structure of the intestinal flora occur in patients with colorectal adenomas, indicating that the association of dysbiosis of the gut microbiota with the occurrence of a pro-oncogenic microenvironment.</p>

Prognosis-relevant factors in primary vaginal carcinoma / 中国肿瘤临床

Objective:To evaluate prognosis factors of primary vaginal cancer. Methods:Data of 80 patients who were hospitalized in the Department of Gynecology and Oncology of Guangxi Tumor Hospital from January 2003 to January 2015 were retrospectively ana-lyzed. All the patients were divided into radiotherapy group (n=49) and surpery group (n=31). Based on radiation mode, the patients with radiotherapy were divided into two-dimensional radiotherapy group (n=29) and three-dimensional radiotherapy (3DRT) group (n=20). Prognosis and complications between two subgroups were compared. Surgical patients were divided into laparoscopic surgery group (n=16) and laparotomy surgery group (n=15) with comparing therapeutic feasibility of video-laparoscopic operation and laparot-omy for primary vaginal carcinoma treatment. Results:Univariate analysis showed that FIGO stage, pathology, tumor size, and extent of vaginal mass involvement were related to prognosis (P<0.05). Multivariate analysis showed that FIGO stage and pathology were in-dependent prognostic factors. Statistical differences of 5-year survival were significant between 2DRT (20.9%) and 3DRT (58.6%) groups (P=0.022). Incidences of urinary tract (14/29, 48.27%) and gastrointestinal symptoms (15/29, 51.72%) in 2DRT group and in 3DRT (3/20,15%;4/20,20%) are different significantly (P<0.05). Hospitalization days of laparotomy surgery group (57.00 ± 41.75) were significantly longer than that of laparoscopic surgery group (29.56 ± 7.30) (P=0.024). Conclusion: Applying laparoscopic surgery and 3DRT improved quality of life without decreasing survival rate of patients with vaginal cancer.

Effect of Down-regulated MUC2 Transcription on Intestinal Permeability in Rats With Acute Necrotizing Pancreatitis / 胃肠病学

Background:Acute pancreatitis can induce intestinal barrier dysfunction in its early phase,which is closely related with the progression and prognosis of the disease. Intestinal mucus layer not only serves as a physical barrier between pathogens and epithelium,but also plays a critical role in the maintenance of intestinal barrier function. Aims:To investigate the expression and role of mucin 2 (MUC2)in injured intestinal barrier in rats with acute necrotizing pancreatitis (ANP). Methods:Forty-two male Sprague-Dawley rats were randomly divided into two groups:the sham operation (SO)group and ANP group,which were induced via a retrograde injection of 3. 5% sodium taurocholate into biliopancreatic duct. Blood,pancreas and colon samples were obtained 6,12 and 24 hours after establishing the ANP model for determination of serum amylase and D-lactate (an indicator of intestinal permeability)and histopathological examination. PAS/ AB staining was used to observe the colon mucus layer and goblet cells,and the expressions of MUC2 and inflammatory cytokines in colonic tissue were detected by real-time PCR. Results:ANP models were successfully established. In ANP group,obvious colonic injury,increased intestinal permeability,thinner colon mucus layer,reduced mucin-containing goblet cells,down-regulated MUC2 mRNA and up-regulated tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β)and interferon-γ (IFN-γ)mRNAs were observed at each time point as compared with those in SO group (P < 0. 05). Spearman correlation coefficient revealed that MUC2 expression was negatively correlated with the intestinal permeability and expression of inflammatory cytokines in ANP group (P < 0. 05). Conclusions:Transcription of MUC2 is significantly down-regulated in colonic tissue of ANP rats,and might be associated with increased intestinal permeability and excessive expression of inflammatory cytokines in early phase of ANP.

Kangle Decoction for erectile dysfunction with liver-qi stagnation and kidney deficiency / 中华男科学杂志

<p><b>Objective</b>To observe the clinical efficacy of Kangle Decoction in the treatment of erectile dysfunction (ED) with liver-qi stagnation and kidney deficiency.</p><p><b>METHODS</b>A total of 79 ED patients with liver-qi stagnation and kidney deficiency were randomly assigned to an experimental group (aged ［36.62±8.05］ yr and with a disease course of ［18.15±6.41］ mo) and a control group (aged ［37.44±8.10］ yr and with a disease course of ［17.51±6.79］ mo), the former treated orally with Kangle Decoction at 0.5 dose bid while the latter with Cialis at 10 mg qd alt, both for 8 weeks. Before treatment, after 4 and 8 weeks of medication, and at 4 weeks after drug withdrawal, we obtained the scores of the patients in the International Index of Erectile Function-5 (IIEF-5), Erectile Dysfunction Inventory of Treatment Satisfaction (EDITS), and Short-Form Psychological and Interpersonal Relationship Scales (SF-PAIRS), and compared the indexes between the two groups of patients.</p><p><b>RESULTS</b>The IIEF-5 score was dramatically increased in both the treatment and control groups after 4 weeks (13.40±2.42 and 16.00±2.68) and 8 weeks of medication (18.60±3.50 and 18.59±3.80) and at 4 weeks after drug withdrawal (17.00±3.05 and 13.95±2.61) as compared with the baseline (10.78±2.28 and 10.77±2.33) (P<0.05 ), even higher in the treatment than in the control group after drug withdrawal (P<0.05 ). The EDITS scores in the treatment and control groups were (28.88±3.31 and 28.90±3.31) after 4 weeks of intervention, (29.68±3.30 and 29.13±3.32) after 8 weeks of intervention, and (29.20±2.92 and 26.82±3.23) at 4 weeks after drug withdrawal, all significantly higher in the former than in the latter group after drug withdrawal (P<0.05 ). The sexual self-confidence score (SSCS), sexual spontaneity score (SSS), and sexual time-concern score (STCS) were all improved in the treatment and control groups after medication as compared with the baseline (P<0.05 ), even higher in the former than in the latter group after drug withdrawal (P<0.05 ).</p><p><b>CONCLUSIONS</b>Kangle Decoction has a definite efficacy in the treatment of ED with liver-qi stagnation and kidney deficiency, with few adverse reactions and long-term post-withdrawal effect, and therefore deserves a wide clinical application.</p>