The relationship between cardiac muscularis propria and clinical outcomes of peroral endoscopic myotomy in achalasia.

BACKGROUND AND AIM: Achalasia patients usually present lower esophageal sphincter thickening, which can impact the expansibility of cardia. We aimed to investigate the effect of cardiac muscularis propria (MP) on perioperative adverse events (AEs) and treatment outcomes of patients treated with peroral endoscopic myotomy (POEM). METHODS: We retrospectively reviewed 114 patients with achalasia undergoing pre-POEM endoscopic ultrasonography (EUS) between May 2013 and November 2019. Cardiac MP thickness was measured using EUS. POEM failure was defined as Eckardt score >3. Risk factors for perioperative AEs and POEM failure were identified. RESULTS: Patients were divided into the thin (n = 52) and the thick group (n = 62) based on the median of cardiac MP thickness (3.0 mm). Perioperative AEs rate of the thin group seemed to be slightly higher than that of the thick group (11.5% vs. 4.8%, P = 0.30). During a median follow-up of 30 months (range 1-77), 100 patients completed follow-up, 16 (16%) of which occurred clinical failure. The clinical outcomes of patients in the thin group were significantly poorer than those patients in the thick group (P = 0.006). Cardiac MP thickness was an independent risk factor for POEM failure (hazard ratio 3.9, P = 0.02; Cox regression), but not the risk factor for perioperative AEs (odds ratio 2.6, P = 0.2; logistic regression). CONCLUSION: Cardiac MP thickness could be a novel predictive factor for POEM failure in patients with achalasia.

IL-22 inactivates hepatic stellate cells via downregulation of the TGF-ß1/Notch signaling pathway.

Interleukin-22 (IL-22) inhibits liver fibrosis by inducing hepatic stellate cell (HSC) senescence, primarily through the activation of signal transducer and activator of transcription 3 signaling. However, whether other signaling pathways are involved remains unknown. The present study assessed the regulatory mechanism between IL­22 and the Notch signaling pathway in vitro. The results revealed that IL­22 had anti­proliferative effects on HSC­T6 cells, and cellular inactivation was reflected by simultaneous inhibition of α­smooth muscle actin, transforming growth factor-ß1 (TGF­ß1), tumor necrosis factor-α and intercellular adhesion molecule 1. Treatment with TGF­ß1 resulted in significant Notch3 upregulation and activation of its downstream effectors Hes family basic helix­loop­helix (bHLH) transcription factor (Hes)-1, Hes­5 and Hes related family BHLH transcription factor with YRPW motif 1. Furthermore, this effect was markedly reversed by further treatment with IL­22, indicating there may be regulatory cascades of IL­22/TGF­ß1/Notch signaling in HSC­T6 cells. The results of the present study demonstrated an inhibitory function of IL­22 towards Notch signaling in hepatic cells, providing evidence that Notch may serve as a novel target for liver fibrosis.

EUS-guided lauromacrogol ablation of insulinomas: a novel treatment.

BACKGROUND AND AIMS: EUS-guided ablation with ethanol has been used to treat insulinoma since 2006 as a minimally invasive alternative for those who are unwilling or unsuitable for surgeries. However, pancreatic fistula, pancreatitis and other adverse effects were found after the procedure in these patients. Herein, we aimed to find a novel feasible injection. METHODS: Seven patients with different chief complaints were diagnosed with insulinoma by symptoms, lab results and pathology results from EUS fine needle aspiration. All the patients refused to have surgeries and were treated by EUS-guided ablation with lauromacrogol. The injection volume was calculated by tumor size. All the patients were followed up by at least 1 month to see if there is any adverse effect. Blood glucose (BG), insulin and C-peptide levels were monitored before and after the procedure. RESULTS: Insulinoma size ranged from 0.76 cm ×0.84 cm to 3.39 cm ×1.84 cm. With a mean injection volume of 1.9 ml (range from 0.9 to 3.9 ml), all the patients showed relief in symptoms after the procedure. During the follow up, their BG, insulin and C-peptide levels went back to normal. None of the patients had any adverse effect. CONCLUSIONS: EUS-guided ablation with lauromacrogol showed good treatment results and received no adverse effect after the procedure. Hence, we consider it as an effective and safe method to treat insulinoma.

Interleukin-22 ameliorates liver fibrosis through miR-200a/beta-catenin.

IL-22 ameliorates liver fibrosis by inhibiting hepatic stellate cells (HSC), and loss of miR-200a is associated with the development of liver fibrosis. The study aimed to investigate the interplay between IL-22 and miR-200a in regulating liver fibrosis in vivo and in vitro. We observed that IL-22 significantly reduced the proliferation of HSC and increased the expression of p-STAT3. ß-catenin was identified as a target gene of miR-200a by luciferase reporter assay, and upregulation of miR-200a significantly attenuated the proliferation of HSC and reduced ß-catenin expression. IL-22 treatment increased expression of miR-200a and decreased expression of ß-catenin in HSC. The expression of p-STAT3 and miR-200a was elevated while ß-catenin was decreased in fibrotic rat liver after IL-22 treatment. Expression levels of ß-catenin and p-STAT3 were inversely correlated in fibrotic rat liver and HSC. Upregulation of ß-catenin suppressed expression of p-STAT3 in HSC. We concluded that IL-22 inhibits HSC activation and ameliorates liver fibrosis through enhancing expression of miR-200a and reducing expression of ß-catenin, suggesting there may be a crosstalk between IL-22/STAT3 and ß-catenin pathway.

A deleterious role for Th9/IL-9 in hepatic fibrogenesis.

T helper 9 (Th9) cells, a recently recognized Th cell subset, are involved in autoimmune diseases. We aimed to investigate the role of Th9/interleukin-9 (IL-9) in the pathogenesis of hepatic fibrosis. Th9 and Th17 cells were quantified in chronic hepatitis B (CHB) patients with hepatic fibrosis, HBV-associated liver cirrhosis (LC) patients and healthy controls (HC). The percentages of Th9 and Th17 cells, concentrations of IL-9 and IL-17, as well as expression of IL-17, TNF-α, IL-6, IL-4, IL-21, TGF-ß1 and IFN-γ were significantly increased in plasma of CHB and LC patients compared with those in HC. Splenic Th9 and Th17 cells, plasma concentrations and liver expression of IL-9 and IL-17A were significantly elevated in mice with hepatic fibrosis compared with controls. Neutralization of IL-9 in mice ameliorated hepatic fibrosis, attenuated the activation of hepatic stellate cells, reduced frequencies of Th9, Th17 and Th1 cells in spleen, and suppressed expression of IL-9, IL-17A, IFN-γ, TGF-ß1, IL-6, IL-4 and TNF-α in plasma and liver respectively. Our data suggest a deleterious role of Th9/IL-9 in increasing hepatic fibrosis and exacerbating disease endpoints, indicating that Th9/IL9 based immunotherapy may be a promising approach for treating hepatic fibrosis.

Interleukin-22 ameliorates liver fibrogenesis by attenuating hepatic stellate cell activation and downregulating the levels of inflammatory cytokines.

AIM: To investigate the effect of interleukin (IL)-22 on hepatic fibrosis in mice and the possible mechanism involved. METHODS: Liver fibrosis was induced in male BALB/c mice by CCl4. Recombinant IL-22 (rmIL-22) was administered intraperitoneally in CCl4-treated mice. Fibrosis was assessed by histology and Masson staining. The activation of hepatic stellate cells (HSCs) was investigated by analysis of α-smooth muscle actin expression. The frequencies of T helper (Th) 22 cells, Th17 cells and Th1 cells, the expression of inflammatory cytokines [IL-22, IL-17A, interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), IL-6, IL-1ß] and transcription factors [aryl hydrocarbon receptor (AHR), RAR-related orphan receptor (RORγt), T-bet] mRNA in the liver were investigated. In addition, the plasma levels of IL-22, IL-17A, IFN-γ, TNF-α, IL-6 and IL-1ß were evaluated. RESULTS: Significant elevations in circulating Th22 cells, Th17 cells, Th1 cells, IL-22, IL-17A, and IFN-γ were observed in the hepatic fibrosis group compared with the control group (P < 0.01). Treatment with rmIL-22 in mice with hepatic fibrosis ameliorated the severity of hepatic fibrosis, which was confirmed by lower hepatic fibrosis pathological scores (P < 0.01). RmIL-22 decreased the frequencies of Th22 cells (6.71% ± 0.97% vs 8.09% ± 0.74%, P < 0.01), Th17 cells (4.34% ± 0.37% vs 5.71% ± 0.24%, P < 0.01), Th1 cells (3.09% ± 0.49% vs 4.91% ± 0.73%, P < 0.01), and the levels of IL-22 (56.23 ± 3.08 vs 70.29 ± 3.01, P < 0.01), IL-17A (30.74 ± 2.77 vs 45.68 ± 2.71, P < 0.01), and IFN-γ (74.78 ± 2.61 vs 124.89 ± 2.82, P < 0.01). Down-regulation of IL-22, IL-17A, IFN-γ, TNF-α, IL-6, IL-1ß, AHR RORγt, and T-bet gene expression in the liver was observed in the rmIL-22 group (P < 0.01). CONCLUSION: The frequencies of Th22, Th17 and Th1 cells are elevated in hepatic fibrosis. RmIL-22 can attenuate HSC activation and down-regulate the levels of inflammatory cytokines, thereby ameliorating liver fibrogenesis.

Th22 cells are associated with hepatocellular carcinoma development and progression.

OBJECTIVE: IL-22-producing CD4(+) T helper cells (Th22 cells) have been identified as major inducers of tissue inflammation and immune responses. Currently, no previous study explored the role of Th22 cells in the pathogenesis of hepatocellular carcinoma (HCC). The study aimed to determine the biological function of Th22 cells and its effector IL-22 in HCC patients. METHODS: Forty-five HCC patients and 19 healthy controls were recruited and their peripheral blood was collected. The fresh HCC tissues, adjacent HCC tissues and ten normal liver tissues were also collected. Flow cytometry analysis was used to determine the frequencies of circulating Th22 cells and Th17 cells. Serum IL-22 levels were tested by enzyme-linked immunosorbent assay (ELISA). Immunohistochemical staining and real-time polymerase chain reaction (PCR) were used to detect IL-22 protein and mRNA in tissues specimens, respectively. RESULTS: Circulating Th22 cells, Th17 cells and serum IL-22 levels were significantly elevated in HCC patients compared with those of healthy controls (P<0.001). Th22 cells were showed to be positively correlated with IL-22 in HCC patients (P<0.05), but not in healthy controls. No significant differences were found in HCC patients with HBeAg positivity or negativity in term of Th22 cells and serum IL-22 levels. The expression of IL-22 protein and mRNA was highest in HCC tissues, followed by adjacent HCC tissues and normal liver tissues. Furthermore, Th22 cells, serum IL-22 levels and IL-22 mRNA were elevated at stage III-IV compared with stage I-II of HCC (P<0.05). CONCLUSIONS: Elevation of circulating Th22 cells and IL-22 may be implicated in the pathogenesis of HCC, and potentially be cellular targets for therapeutic intervention.

Association of interleukin-10 polymorphisms with risk of irritable bowel syndrome: a meta-analysis.

AIM: To clarify the current understanding of the association between interleukin-10 (IL-10) polymorphisms and the risk of irritable bowel syndrome (IBS). METHODS: We searched for studies in any language recorded in PubMed, Embase and Cochrane library before August 2013. The associations under allele contrast model, codominant model, dominant model, and recessive model were analyzed. The strengths of the association between IL-10 polymorphisms and IBS risk were estimated using odds ratios (OR) with 95% confidence interval (CI). Fixed effects model was used to pool the result if the test of heterogeneity was not significant, otherwise the random-effect model was selected. RESULTS: Eight case-control studies analyzing three single-nucleotide polymorphisms rs1800870 (-1082 A/G), rs1800871 (-819C/T), and rs1800872 (-592A/C) of the IL-10 gene, which involved 928 cases and 1363 controls, were eligible for our analysis. The results showed that rs1800870 polymorphisms were associated with a decreased risk of IBS (GG+GA vs AA: OR = 0.80, 95%CI: 0.66-0.96), (AA+GA vs GG: OR = 0.68, 95%CI: 0.52-0.90). Subgroup analysis revealed such association only existed in Caucasian ethnicity (AA+GA vs GG, OR = 0.70, 95%CI: 0.55-0.89). The rs1800872 polymorphisms were associated with an increased risk of IBS in Asian ethnicity (CC vs GG: OR = 1.29, 95%CI: 1.01-1.16). There were no associations between rs1800871 polymorphisms and the IBS risk. CONCLUSION: The results suggest that IL-10 rs1800870 confers susceptibility to the risk of IBS in Caucasian ethnicity, and the rs1800872 may associate with IBS risk in Asians. However, no significant associations are found between rs1800871 and IBS risk.

Alterations of maternal serum and placental follistatin-like 3 and myostatin in pre-eclampsia.

AIM: To clarify the alterations of myostatin, a member of the transforming growth factor-ß superfamily, and follistatin-like 3 (FSTL3), a binding protein for myostatin, in pre-eclamptic women. METHODS: Samples of blood and placenta were collected from 40 pre-eclamptic women and 40 controls. The serum level and placental expression of FSTL3 and myostatin were determined with enzyme-linked immunosorbent assay, real-time polymerized chain reaction and western blotting. RESULTS: The serum levels of myostatin and FSTL3 were significantly higher in pre-eclamptic women than in the controls (P < 0.001 for both). Placental expression of myostatin and FSTL3 were also significantly increased in the pre-eclamptic placenta compared with that of the controls (P < 0.001 for both); however, there were no significant differences in myostatin or FSTL3 in either the maternal serum or the placenta in women with mild or severe pre-eclampsia (P > 0.05 for both). CONCLUSION: The serum levels and placental expression of myostatin and FSTL3 are elevated in pre-eclampsia, suggesting the role of myostatin and its binding protein in pre-eclampsia.

[Effects of PRL-3 siRNA on the migration of endometriotic stromal cells].

OBJECTIVE: To evaluate the effects of PRL-3 siRNA (small interfering RNA) on the migration of endometriotic stromal cells. METHODS: Primary endometriotic stromal cells were cultured in vitro. Then PRL-3 (phosphatase of regenerating liver-3) siRNA was transfected to silence the PRL-3 gene. And the PRL-3 protein expression was analyzed by Western blot. The changes of cell migration were detected by cell pseudopod formation, scratch test and transwell cell migration test. RESULTS: The expression of PRL-3 protein significantly decreased in the experimental group versus two other control groups (P < 0.05). The formation of cell pseudopod was much less in experimental group than that in control groups. Within the same time, the number of migration cells was 0.87 ± 0.21 in experimental group. It was less than 1.75 ± 0.28 in blank control group and 1.63 ± 0.39 in negative control group (P < 0.05). CONCLUSION: PRL-3 siRNA can down-regulate the PRL-3 gene and decrease the migratory capacity of endometriotic stromal cells. And PRL-3 may be a promising target in the therapeutics of endometriosis.

[Association between susceptibility of endometriosis and the gene polymorphism of tumor necrosis factor].

OBJECTIVE: To evaluate the association between the susceptibility of endometriosis (EMS) and the gene polymorphism of tumor necrosis factor (TNF) in Chinese Han population. METHODS: Peripheral blood samples were collected from 76 EMS patients and 87 normal controls. PCR-RFLP was used to detect the single base change polymorphism in both TNFalpha gene and TNFbeta gene. RESULTS: The allele frequencies of the TNFalpha polymorphism were 0.9474 and 0.9253 in the EMS patients and the control group respectively, and the TNFbeta polymorphism were 0.0526 and 0.0747 respectively. The allele frequencies of the TNFbeta1 polymorphism were 0.4605 and 0.5115 in the EMS patients and the control group, and the TNFbeta2 polymorphism were 0.5395 and 0.4885. No difference in the gene and genotype frequencies of TNFalpha and TNFbeta was noted in the EMS patients and normal controls (all P > 0.05). CONCLUSION: TNF gene polymorphism has no effect on EMS in Chinese Han population. Ethnic difference may exist in TNF allele frequency.

Serum adiponectin, leptin and soluble leptin receptor in pre-eclampsia.

OBJECTIVES: To delineate the changes in serum levels of adiponectin, leptin and soluble leptin receptor, and in the free leptin index in women with pre-eclampsia. METHODS: Blood samples were collected from 38 pregnant women with pre-eclampsia and 42 normotensive pregnant women as controls. Serum concentrations of adiponectin, leptin and soluble leptin receptor were determined by enzyme-link immunosorbent assay and the free leptin index was calculated as the ratio of serum leptin to soluble leptin receptor for each sample. RESULTS: No significant differences were observed between the groups regarding maternal age, gestational age and body mass index. Women with pre-eclampsia had significantly higher levels of serum adiponectin and leptin, and a higher free leptin index than controls (P<0.01, P<0.001 and P<0.001, respectively). There were no significant differences between the two groups in serum levels of soluble leptin receptor (P>0.05). CONCLUSIONS: The study demonstrated elevated serum levels of adiponectin and leptin as well as a higher free leptin index in women with pre-eclampsia, suggesting these as important factors contributing to this complication of pregnancy.

[Changes of serum cardiotrophin I and cystatin C in preeclampsia and clinical significance thereof].

OBJECTIVE: To investigate the changes and of serum cardiotrophin I (cTN I) and cystatin C in preeclampsia clinical significance thereof. METHODS: Peripheral blood samples were collected from 40 women with preeclampsia, aged 29 (18 - 39), with the medium gestational age of 35 weeks, and 40 age- and gestational age-matched controls. Serum cTN I and cystatin C were determined with ELISA and serum levels of creatinine kinase, creatinine, and uric acid were assayed by automatic biochemical analyzer. Comparison was made between the 2 groups. RESULTS: The serum cTn I of the preeclampsia group was 0.59 microg/L +/- 0.42 microg/L, significantly higher than that of the control group (0.09 microg/L +/- 0.22 microg/L P < 0.001), and the serum cystatin C of the preeclampsia group was 1.64 mg/L +/- 0.47 mg/L, significantly higher than that of the control group (1.08 mg/L +/- 0.19 mg/L, P < 0.001). And the serum cardiotrophin I and cystatin C concentrations of the patients with severe preeclampsia were 0.66 microg/L +/- 0.40 microg/L and 1.72 mg/L +/- 0.46 mg/L respectively, both significantly higher than those of the patients with mild preeclampsia (0.17 microg/L +/- 0.26 microg/L and 1.21 mg/L +/- 0.37 mg/L respectively, P < 0.001 and 0.05 respectively). CONCLUSION: Serum cTn I and cystatin C levels are elevated in preeclampsia and the elevation is associated with the severity of preeclampsia, suggesting that serum concentrations of cTn I and cystatin C are useful markers in the early diagnosis of cardiac and renal injury in patients of preeclampsia.