Role of hydrogen sulfide in portal hypertension and esophagogastric junction vascular disease

AIM:To investigate the association between endogenous hydrogen sulfide(H2S)and portal hypertension as well as its effect on vascular smooth muscle cells.METHODS:Portal hypertension patients were categorized by Child-Pugh score based on bilirubin and albumin levels,prothrombin time,ascites and hepatic encephalopathy.Plasma H2S concentrations and portal vein diameters(PVDs)were compared between portal hypertension patients and control participants,as well as between portal hypertension patients with varying degrees of severity.In addition,we established a rabbit hepatic schistosomiasis portal hypertension(SPH)model and analyzed liver morphology,fibrosis grade,plasma and liver tissue H2S concentrations,as well as cystathionineγ-lyase(CSE)activity and phosphorylated extracellular signal-regulated kinase(pERK)1/2,B cell lymphoma(Bcl)-2 and Bcl-XL expression in portal vein smooth muscle cells,in addition to their H2S-induced apoptosis rates.RESULTS:In portal hypertension patients,endogenous H2S levels were significantly lower than those in healthy controls.The more severe the disease was,the lower were the H2S plasma levels,which were inversely correlated with PVD and Child-Pugh score.Liver tissue H2S concentrations and CSE expression were significantly lower in the SPH rabbit livers compared with the control animals,starting at 3 wk,whereas pERK 1/2expressions gradually increased 12-20 wk after SPH model establishment.In portal vein smooth muscle cells,increasing H2S levels led to increased apoptosis,while Bcl-2 and Bcl-XL expression decreased.CONCLUSION:H2S prevents vascular restructuring caused by excessive proliferation of smooth muscle cells via apoptosis induction,which helps to maintain normal vascular structures.

Replication-incompetent Adenovirus Vector-mediated MDA-7/IL-24 Selectively Induces Growth Suppression and Apoptosis of Hepatoma Cell Line SMMC-7721

In order to investigate the effect of replication-incompetent adenovirus vector expressing MDA-7/IL-24 on tumor growth and apoptosis of human hepatocellular carcinoma （HCC） cell line SMMC-7721 and normal liver cell line L02, the recombinant replication-incompetent Ad.mda-7 virus vector was constructed and infected into the HCC cell line SMMC-7721 and normal liver cell line L02. RT-PCR was performed to examine the expression of MDA-7 mRNA. The concentrations of MDA-7/IL-4 in culture superuatants were determined by using ELISA. MTT and Hoechst staining assay were applied to observe the inhibitory and killing effects of MDA-7 on the HCC cells. By using flow cytometry, the apoptosis, cell cycle and proliferation of SMMC-7721 and L02 cells were measured. The results showed recombinant replication-incompetent virus expressing MDA-7/IL-24 was constructed successfully, and RT-PCR revealed that it could mediate the high expression of the exogenous gene MDA-7/IL-24 in SMMC-7721 and L02 cells. The expression of MDA-7/IL-24 proteins in the culture superuatant was detectable by ELISA. Ad.mda-7 infection induced apoptosis and growth suppression in SMMC-7721 cells and an increased percentage of HCC cells in the GyM phase of the cell cycle, but not in L02 cells. It was concluded that mda-7/IL-24 gene, mediated with replication-incompetent adenovirus vector, could selectively induce growth suppression and apoptosis in HCC cell line SMMC-7721 but without any toxic side-effect on normal liver line L02.

Targeted bile acids metabolomics in cholesterol gallbladder polyps and gallstones: From analytical method development towards application to clinical samples

Bile acids(BAs)are synthesized by the liver from cholesterol through several complementary pathways and aberrant cholesterol metabolism plays pivotal roles in the pathogeneses of cholesterol gallbladder polyps(CGP)and cholesterol gallstones(CGS).To date,there is neither systematic study on BAs profile of CGP or CGS,nor the relationship between them.To explore the metabolomics profile of plasma BAs in healthy volunteers,CGP and CGS patients,an ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method was developed and validated for simultaneous determination of 42 free and conjugated BAs in human plasma.The developed method was sensitive and reproducible to be applied for the quantification of BAs in the investigation of plasma samples.The results show that,compared to healthy volunteers,CGP and CGS were both characterized by the significant decrease in plasma BAs pool size,furthermore CGP and CGS shared aberrant BAs metabolic characteristics.Chenodeoxycholic acid,glycochenodeoxycholic acid,l-muricholic acid,deoxycholic acid,and 7-ketolithocholic acid were shared potential markers of these two cholesterol gallbladder diseases.Subsequent analysis showed that clinical characteristics including cysteine,ornithine and body mass index might be closely related to metabolisms of certain BA modules.This work provides metabolomic information for the study of gallbladder diseases and analytical methodologies for clinical target analysis and efficacy evaluation related to BAs in medical institutions.

Radical nerve dissection for the carcinoma of head of pancreas: report of 30 cases

Objective： To explore the clinical value of the radical nerve dissection （RND） for the carcinoma of head of pancreas （CHP）. Methods： The clinical and pathological data of 30 CHP patients who underwent RND in our hospital were retrospectively analyzed, with an attempt to explore the safety and short-term efficacy of this procedure. Results： Among these 30 patients, the operative time was （351±61） rain, the intra-operative blood loss was 350 （range, 300-600） mL, and the grades B and C pancreatic fistula was 23.33%. During the follow-up （range, 2-30 months; median： 17 months）, the 1-year survival rate was 63.33% and the 1-year disease-free survival rate was 56.67%. Among the 23 patients （76.66%） with positive extra-pancreatic perineural invasion （PNI）, the 1-year case- fatality rate was 34.78%, which was not significantly different from that （28.57%） of patients with negative PNI （P=0.760）. Conclusions： Our results suggested potential advantages of RND in the fields of surgery-associated risk and prognosis compared with the Whipple operation in the treatment of CHP. Due to the low sample size of this study, further well-designed research of large sample size is needed.

Different Optical Properties between Human Hepatocellular Carcinoma Tissues and Non-tumorous Hepatic Tissues In Vitro

There has been an ongoing search for clinically acceptable methods for the accurate,efficient and simple diagnosis and prognosis of hepatocellular carcinoma（HCC）.Optical spectroscopy is a technique with potential clinical applications to diagnose cancer diseases.The purpose of this study was to obtain the optical properties of HCC tissues and non-tumorous hepatic tissues and identify the difference between them.A total of 55 tissue samples（HCC tissue,n=38;non-tumorous hepatic tissue,n=17） were surgically resected from patients with HCC.The optical parameters were measured in 10-nm steps using single-integrating-sphere system in the wavelength range of 400 to 1800 nm.It was found that the optical properties and their differences varied with the wavelength for the HCC tissue and the non-tumorous hepatic tissue in the entire wavelength range of research.The absorption coefficient of the HCC tissue（1.48±0.99,1.46±0.88,0.86±0.61,2.15±0.53,0.54±0.10,0.79±0.15 mm-1） was significantly lower than that of the non-tumorous hepatic tissue（2.79±1.73,3.13±1.47,3.06±2.79,2.57±0.55,0.62±0.10,0.93±0.16 mm-1） at wavelengths of 400,410,450,1450,1660 and 1800 nm,respectively（P0.05）.The reduced scattering coefficient of HCC tissue（5.28±1.70,4.91±1.54,1.26±0.35 mm-1） and non-tumorous hepatic tissue（8.14±3.70,9.27±3.08,2.55±0.57 mm-1） was significantly different at 460,500 and 1800 nm respectively（P0.05）.These results show different pathologic liver tissues have different optical properties.It provides a better understanding of the relationship between optical parameters and physiological characteristics in human liver tissues.And it would be very useful for developing a non-invasive,real-time,simple and efficient way for medical management of HCC in the future.

Protective Effects of Hydrogen Sulfide on Portal Hypertensive Vasculopathy in Rabbits by Activating AKT-NF-κB Pathway

The role of hydrogen sulfide（H;S） in portal hypertension（PH）-induced esophagus-gastric junction vascular lesions in rabbits was observed. The rabbit PH models were established. The animals were randomly divided into the following groups: normal, PH, PH+sodium hydrosulfide（PH+S）, PH+propargylglycine（PH+PPG）. The plasma H;S levels, apoptosis of esophageal-gastric junction vascular smooth muscle cells, and the expression of nuclear transcription factor-κB（NF-κB）, p-AKT, IκBa and Bcl-2 were detected. The cystathionine γ lyase（cystathionine-gamma-splitting enzyme, CSE） in the junction vascular tissue was measured. The results showed that the plasma H;S levels and the CSE expression levels had statistically significant difference among different groups（P<0.05）. As compared with PH group, plasma H;S levels were declined obviously（11.9±4.2 vs. 20.6±4.5, P<0.05）, and CSE expression levels in the junction vascular tissue were notably reduced（1.7±0.6 vs. 2.8±0.8, P<0.05）, apoptosis rate of vascular smooth muscle cells per unit area was significantly decreased（0.10±0.15 vs. 0.24±0.07, P<0.05）, and the expression levels of p-AKT and NF-κB were significantly decreased（2.31±0.33 vs. 3.04±0.38, P<0.05; 0.33±0.17 vs. 0.51±0.23, P<0.05）, however, IκBa and Bcl-2 expression increased obviously（5.57±0.17 vs. 3.67±0.13, P<0.05; 0.79±0.29 vs. 0.44±0.36, P<0.05） in PH+PPG group. As compared with PH group, H;S levels were notably increased（32.7±7.3 vs. 20.6±4.5, P<0.05）, the CSE levels in the junction vascular tissue were significantly increased（6.3±0.7 vs. 2.8±0.8, P<0.05）, apoptosis rate of vascular smooth muscle cells per unit area was significantly increased（0.35±0.14 vs. 0.24±0.07, P<0.05）, and the expression levels of p-AKT and NF-κB were significantly increased（4.29±0.49 vs. 3.04±0.38, P<0.05; 0.77±0.27 vs. 0.51±0.23, P<0.05）, yet IκBa and Bcl-2 expression decreased significantly（3.23±0.24 vs. 3.67±0.13, P<0.05; 0.31±0.23 vs. 0.48±0.34, P<0.05） in PH+S group. It is concluded that esophagus-gastric junction vascular lesions happen under PH, and apoptosis of smooth muscle cells is declined. H;S can activate NF-κB by the p-AKT pathway, leading to the down-regulation of Bcl-2, eventually stimulating apoptosis of vascular smooth muscle cells, easing PH. H;S/CSE system may play an important role in remission of PH via the AKT-NF-κB pathway.

Identification of two migratory colon ILC2 populations differentially expressing IL-17A and IL-5/IL-13

Group 2 innate lymphoid cells(ILC2s)play important tissue resident roles in anti-parasite immunity,allergic immune response,tissue homeostasis,and tumor immunity.ILC2s are considered tissue resident cells with little proliferation at steady state.Recent studies have shown that a subset of small intestinal ILC2s could leave their residing tissues,circulate and migrate to different organs,including lung,liver,mesenteric LN and spleen,upon activation.However,it remains unknown whether other ILC populations with migratory behavior exist.In this study,we find two major colon ILC2 populations with potential to migrate to the lung in response to IL-25 stimulation.One subset expresses IL-17A and resembles inflammatory ILC2s(iILC2s)but lacks CD27 expression,whereas the other expresses CD27 but not IL-17A.In addition,the IL-17A^(+)ILC2s express lower levels of CD127,CD25,and ST2 than CD27^(+)ILC2s,which express higher levels of IL-5 and IL-13.Surprisingly,we found that both colon ILC2 populations still maintained their colonic features of preferential expression of IL-17A and CD27,IL-5/IL-13,respectively.Together,our study identifies two migratory colon ILC2 subsets with unique surface markers and cytokine profiles which are critical in regulating lung and colon immunity and homeostasis.

Prevention and treatment of pancreatic fistula after pancreatic body and tail resection: current status and future directions

Postoperative pancreatic fistula (POPF) is the most common and critical complication after pancreatic body and tail resection.How to effectively reduce the occurrence of pancreatic fistula and conduct timely treatment thereafter is an urgent clinical issue to be solved.Recent research standardized the definition of pancreatic fistula and stressed the correlation between POPF classification and patient prognosis.According to the literature,identification of the risk factors for pancreatic fistula contributed to lowering the rate of the complication.Appropriate management of the pancreatic stump and perioperative treatment are of great significance to reduce the rate of POPF in clinical practice.After the occurrence of POPF,the treatment of choice should be determined according to the classification of the pancreatic fistula.However,despite the progress and promising treatment approaches,POPF remains to be a clinical issue that warrants further studies in the future.

Melanoma differentiation-associated gene-7, MDA-7/IL-24, selectively induces growth suppression, apoptosis in human hepatocellular carcinoma cell line HepG2 by replication-incompetent adenovirus vector

瞄准：调查在人的肝细胞在肿瘤生长和 apoptosis 上表示 MDA-7/IL-24 的复制无能的侵入人体气管粘膜的病菌向量的效果癌(HCC ) 房间线 HepG2 和正常的肝房间衬里 L02。方法：我们构造了 recombinant 复制无能的 Ad.mda-7 病毒向量并且感染了它进人的 HCC 房间线 HepG2 和正常的肝房间线 L02。RT-PCR 被执行检测在房间表示的 mRNA。被 ELISA 用来在文化上层清液检测 MDA-7/IL-24 蛋白质表示。Ad.mda-7 导致的 apoptosis 的效果被 Hoechst 与染色的 Annexin-V 和 PI 染色和流动血细胞计数试金证实。MTT 试金被用来决定 HepG2 细胞的生长抑制，并且房间周期和 hypodiploidy 分析被流动血细胞计数执行。结果：Recombinant 复制有缺陷者病毒表示 MDA-7/IL-24 成功地被构造。RT-PCR 证明 Ad.mda-7 能调停进 HepG2 和 L02 的外长的基因 MDA-7/IL-24 的表示。在上层清液的 MDA-7/IL-24 蛋白质的集中分别地在 Ad.mda-7-infected L02 和 HepG2 房间是 130 pg/mL 和 110 pg/mL。 Ad.mda-7 感染显然导致了 apoptosis (从2.60%+/-0.72%到33.6%+/-13.2%， P=0.00012 )并且在 HepG2 (抑制比率IR=68%)的生长抑制和在房间的 G2/M 阶段的特定的癌症房间类型的百分比的增加骑车(从6.44%～32.29%， P【0.01 )，然而并非在 L02 房间。结论：这些结果有选择地在人的肝细胞癌细胞线 HepG2 与复制无能的侵入人体气管粘膜的病菌向量由 mda-7/IL-24 基因证实 apoptosis 和生长抑制的正式就职。

Oncolytic adenovirus SG600-IL24 selectively kills hepatocellular carcinoma cell lines

AIM: To investigate the effect of oncolytic adenovirus SG600-IL24 and replication-incompetent adenovirus Ad.IL-24 on hepatocellular carcinoma (HCC) cell lines and normal liver cell line. METHODS: HCC cell lines (HepG2, Hep3B and MHCC97L) and normal liver cell line (L02) with a different p53 status were infected with SG600-IL24 and Ad.IL-24, respectively. Melanoma differentiation-associated (MDA)-7/interleukin (IL)-24 mRNA and protein expressions in infected cells were detected by reverse transcription-polymerase chain reaction (RT-PCR), enzymelinked immunosorbent assay (ELISA), and Western blotting, respectively. Apoptosis of HCC cells and normal liver cells was detected by cytometric assay with Hoechst33258 staining. 3-(4,5-dimethylthiazol-2-yl)-2,5diphenyltetrazolium bromide (MTT) assay was used to investigate proliferation of HCC cells and normal liver cells, and cell cycle was assayed by flow cytometry. RESULTS: RT-PCR, ELISA and Western blotting showed that the exogenous MDA-7/IL-24 gene was highly expressed in cells infected with SG600-IL24. MTT indicated that SG600-IL24 could suppress the growth of HepG2, Hep3B, MHCC97L, with an inhibition rate of 75% ± 2.5%, 85% ± 2.0%, 72% ± 1.8%, respectively (P < 0.01), promote the apoptosis of HepG2, Hep3B, MHCC97L, with an apoptosis rate of 56.59% ± 4.0%, 78.36% ± 3.5%, 43.39% ± 2.5%, respectively (P < 0.01), and block the HCC cell lines in the G2/M phase with a blocking rate of 35.4% ± 4.2%, 47.3% ± 6.2%, 42% ± 5.0%, respectively (P < 0.01) but not the normal liver cell line in a p53-independent manner. CONCLUSION: SG600-IL24 can selectively suppress the proliferation and apoptosis of HCC cell lines in vitro but not normal liver cell line L02 in a p53-independent manner. Compared with Ad.IL-24, SG600-IL24 can significantly enhance the antitumor activity in HCC cell lines.

Disease control with sunitinib in advanced intrahepatic cholangiocarcinoma resistant to gemcitabine-oxaliplatin chemotherapy

Advanced cholangiocarcinoma is associated with poor prognostic survival and has limited therapeutic options available at present. The importance of angiogenesis and expression of pro-angiogenic factors in intrahepatic forms of cholangiocarcinoma suggest that therapies targeting angiogenesis might be useful for the treatment of this disease. Here we report three cases of patients with advanced intrahepatic cholangiocarcinoma progressive after standard chemotherapy and treated with sunitinib 50 mg/d in 6-wk cycles of 4 wk on treatment followed by 2 wk off treatment(Schedule 4/2). In all three patients, sunitinib treatment was associated with a sustained disease control superior to 4 mo, patients achieving either a partial response or stable disease. A reduction in tumor size and density was observed in all cases, suggesting tumor necrosis as a result of sunitinib treatment in these patients. In addition, sunitinib was generally well tolerated and the occurrence of side effects was managed with standard medical interventions, as required. Our results suggest that sunitinib therapy maybe associated with favorable outcomes and tolerability in patients with advanced cholangiocarcinoma. Those observations contributed to launch a prospective phase Ⅱ multicenter trial investigating sunitinib in advanced intrahepatic cholangiocarcinoma(SUN-CK study; NCT01718327).

Application of bioactive hydrogels combined with dental pulp stem cells for the repair of large gap peripheral nerve injuries

Due to the limitations in autogenous nerve grafting or Schwann cell transplantation,large gap peripheral nerve injuries require a bridging strategy supported by nerve conduit.Cell based therapies provide a novel treatment for peripheral nerve injuries.In this study,we first experimented an optimal scaffold material synthesis protocol,from where we selected the 10%GFD formula(10%GelMA hydrogel,recombinant human basic fibroblast growth factor and dental pulp stem cells(DPSCs))to fill a cellulose/soy protein isolate composite membrane(CSM)tube to construct a third generation of nerve regeneration conduit,CSM-GFD.Then this CSM-GFD conduit was applied to repair a 15-mm long defect of sciatic nerve in a rat model.After 12 week post implant surgery,at histologic level,we found CSM-GFD conduit could regenerate nerve tissue like neuron and Schwann like nerve cells and myelinated nerve fibers.At physical level,CSM-GFD achieved functional recovery assessed by a sciatic functional index study.In both levels,CSM-GFD performed like what gold standard,the nerve autograft,could do.Further,we unveiled that almost all newly formed nerve tissue at defect site was originated from the direct differentiation of exogeneous DPSCs in CSM-GFD.In conclusion,we claimed that this third-generation nerve regeneration conduit,CSM-GFD,could be a promising tissue engineering approach to replace the conventional nerve autograft to treat the large gap defect in peripheral nerve injuries.

Corrigendum to‘Application of bioactive hydrogels combined with dental pulp stem cells for the repair of large gap peripheral nerve injuries’[Bioactive Mater.6(3)(2021)638-654]

The authors regret that the author(A.H.)was removed as an agreement could not be reached between the two affiliations of the author.The authors would like to apologise for any inconvenience caused.