Anti-tumour effect of Huangqin Decoction on colorectal cancer mice through microbial butyrate mediated PI3K/Akt pathway suppression.

Introduction. Huangqin Decoction (HQD), a Chinese herbal formula, is widely used for various diseases, including colorectal cancer (CRC).Hypothesis/Gap Statement. We proposed that microbial butyrate mediated PI3K/Akt pathway suppression might involve the anti-cancer effect of HQD.Aim. This study aimed to evaluate the potential mechanism of HQD against CRC.Methodology. An azoxymethane plus dextran sulphate sodium induced CRC mouse model was used, and the intestinal flora and faecal short-chain fatty acid changes were detected, respectively, after HQD administration with 16S rRNA sequencing and gas chromatography coupled with mass spectrometry. Disease activity index, colon length and levels of inflammatory cytokines were measured to evaluate the effect of HQD on intestinal inflammation. Tumour size, number and histopathology were assessed to reflect the impact of HQD on tumour burden. Apoptosis and PI3K/Akt pathway activity were measured by TUNEL staining and Western-blotting. In vitro, the effects of sodium butyrate (NaB) on the viability of CRC cell lines were detected by the Cell-counting Kit-8. The apoptotic cells were determined by TUNEL staining. Cell migration and invasion were assessed by wound healing assay and Transwell assay, respectively. Western-blotting and immunofluorescent staining were used to test the activity of PI3K/Akt pathway.Results. Animal study showed that HQD could improve the gut dysbiosis, increase the abundance of Clostridium and the level of faecal butyric acid. Then, we found that HQD could attenuate colitis, reduce tumour burden, promote cell apoptosis and suppress PI3K/Akt pathway activity in CRC mice. In vitro experiment revealed that NaB treatment could inhibit cell growth, migration and invasion in CRC cell lines. Additionally, NaB enhanced cellular apoptosis, and reduced phosphorylated PI3K and Akt expressions. Interestingly, addition of 740Y-P, an agonist of PI3K, reversed the NaB effects on CRC cells.Conclusion. Overall, in this study, we revealed that HQD could induce apoptosis through microbial butyrate mediated PI3K/Akt inhibition and perform anti-CRC activity.

FoxM1 is associated with metastasis in colorectal cancer through induction of the epithelial-mesenchymal transition.

The aim of the present study was to investigate the role of forkhead box M1 (FoxM1) in epithelial-mesenchymal transition (EMT) and metastasis in colorectal cancer (CRC). Immunohistochemical assays were performed to detect FoxM1 and epithelial (E-) cadherin protein expression in 92 CRC, 61 colonic adenoma and 32 wild-type colonic tissue samples. Quantitative polymerase chain reaction (qPCR) assays were performed to determine the expression levels of FoxM1 and E-cadherin mRNAs in 30 CRC and adjacent normal mucosal tissues. RNA interference was used to knock down endogenous FoxM1 expression in CRC cell lines, and the migratory and invasive capacity of the CRC cells was analyzed. The expression of FoxM1, E-cadherin and neuronal (N-) cadherin in the CRC cell lines was evaluated using qPCR and Western blot analysis. The relative expression levels of FoxM1 mRNA and protein were significantly increased in the CRC tissues compared with those in the colonic adenoma and wild-type mucosal tissue samples (P<0.01). In contrast, the relative expression levels of E-cadherin mRNA and protein were significantly decreased in the CRC tissues compared with in the colonic adenoma and normal mucosal tissues (P<0.01). FoxM1 overexpression and decreased E-cadherin expression were significantly associated with poor colonic tissue differentiation, lymph node metastasis and an advanced tumor-node-metastasis stage. Additionally, the increased expression of FoxM1 was associated with a decrease in E-cadherin expression (P<0.01). Furthermore, RNA interference-mediated FoxM1 knockdown significantly inhibited the proliferation, migration and invasion of CRC cells. Downregulation of FoxM1 expression significantly increased E-cadherin expression and decreased N-cadherin expression. The results of the present study suggest that FoxM1 overexpression in tumor tissues is significantly associated with metastasis in CRC through the induction of EMT.

Hyperechoic demarcation line between a tumor and the muscularis propria layer as a marker for deciding the endoscopic treatment of gastric submucosal tumor.

Minimally invasive endoscopic resection has been rapidly adopted as a new technique for treating patients with gastric submucosal tumors (SMTs) originating in the muscularis propria (MP) layer. This study was conducted to evaluate the information obtained from endoscopic ultrasonography (EUS) to determine the appropriate endoscopic dissection method for treating SMTs originating in the MP layer. Between February 2014 and May 2016, a total of 50 patients with gastric SMTs originating in the MP layer were enrolled in this study. The clinical features of the patients and their endoscopic, EUS, and histopathologic findings, as well as their postoperative follow-up data, were analyzed in this retrospective study. The mean age of the patients was (55.0±10.2) years, and the male/female ratio was 17:33. Endoscopic submucosal dissection (ESD) was performed on 43 patients and an endoscopic full-thickness resection (EFR) was performed on seven patients. The most frequent location for an SMT was in the upper body region of the stomach (n=16), and the most common pathological diagnosis was a gastrointestinal stromal tumor (GIST) (n=32). The overall rates for complete resection were 95.3% (41/43) and 100.0% (7/7) when the SMTs were treated by ESD and EFR, respectively. The presence of a complete tumor capsule was significantly associated with a complete resection (P=0.001). Of the cases treated by ESD, nine patients developed perforation, one of whom required laparoscopic surgery. The remaining patients were closed with clips or purse-string sutures. The presence of an MP2-type tumor (P=0.018) and a wide connection with the MP layer (P=0.044) were significantly associated with perforation. A preoperative evaluation of the integrity and the location of a tumor capsule and the length of the tumor connection with the MP layer by EUS can improve the complete resection rate and reduce the occurrence of intraoperative complications. Tumors with a complete capsule originating from the superficial MP layer or with a narrow connection with the MP layer are appropriate candidates for treatment by ESD.

Successful treatment of refractory gastric antral vascular ectasia by distal gastrectomy: a case report.

Gastric antral vascular ectasia (GAVE) is an uncommon and often neglected cause of gastric hemorrhage. The treatments for GAVE include surgery, endoscopy and medical therapies. Here, we report an unusual case of GAVE. A 72-year-old man with a three-month history of recurrent melena was diagnosed with GAVE. Endoscopy revealed the classical "watermelon stomach" appearance of GAVE and complete pyloric involvement. Melena reoccurred three days after argon plasma coagulation treatment, and the level of hemoglobin dropped to 47 g/L. The patient was then successfully treated with distal gastrectomy with Billroth II anastomosis. We propose that surgery should be considered as an effective option for GAVE patients with extensive and severe lesions upon deterioration of general conditions and hemodynamic instability.

Expression of galectin-1 in carcinoma-associated fibroblasts promotes gastric cancer cell invasion through upregulation of integrin ß1.

Increased expression of galectin-1 (Gal-1) in carcinoma-associated fibroblasts (CAFs) has been reported to correlate with progression and prognosis in many cancers. However, rarely have reports sought to determine whether high Gal-1 expression in CAFs in gastric cancer is involved in the tumor process, and the specific mechanism by which it promotes the evolution of gastric cancer is still unknown. In this study, we cultured gastric cancer CAFs, which showed strong expression of Gal-1, and established a co-culture system of CAFs with gastric cancer cells. Specific siRNA and in vitro migration and invasion assays were used to explore the effects of the interaction between Gal-1 expression of CAFs and gastric cancer cells on cell migration and invasion. We found that the overexpression of Gal-1 in CAFs enhanced gastric cancer cell migration and invasion, and these stimulatory effects could be blocked by specific siRNA which reduced the Gal-1 expression level. A set of cancer invasion-associated genes were then chosen to identify the possible mechanism of Gal-1-induced cell invasion. Among these genes, integrin ß1 expression in cancer cells was considered to be associated with Gal-1 expression. Pre-blocking of the integrin ß1 expression in gastric cancer cells with siRNA could interrupt the invasion-promoting effect of CAFs with high Gal-1 expression. Furthermore, immunohistochemical assay confirmed a positive correlation between Gal-1 and integrin ß1 expression. Our results showed that high expression of Gal-1 in CAFs might facilitate gastric cancer cell migration and invasion by upregulating integrin ß1 expression in gastric cancer.

Subcapsular hepatic haematoma after endoscopic retrograde cholangiopancreatography: an unusual case.

Subcapsular hepatic haematoma is a rare complication of endoscopic retrograde cholangiopancreatography (ERCP), and there are few reports about this unusual complication worldwide. The primary symptom of most cases reported in the literature is abdominal pain. We report an unusual case with the primary symptom of fever. A 56-year-old man who had a six-month history of recurrent episodes of upper abdominal pain was diagnosed with a common bile duct stone by magnetic resonance cholangiopancreatography. Endoscopic biliary sphincterotomy was performed, and stones from the common bile duct were successfully extracted with a basket. The patient had a persistent fever after ERCP, and treatment with intravenous antibiotics was unsuccessful. Computed tomography showed a 13 cm × 6 cm subcapsular hepatic haematoma filled with air and liquid on the surface of the right hepatic lobe. The patient was successfully treated with peritoneal drainage under B-ultra guidance. Subcapsular liver haematoma should be considered when hard-to- explain symptoms persist in the early period after ERCP. Percutaneous drainage is an effective treatment.

The accumulating appreciation of endoscopic submucosal dissection in the treatment of gastrointestinal neoplasms: preliminary experience in local eastern China.

BACKGROUND/AIMS: Endoscopic submucosal dissection (ESD), an emerging technique originated from Japan, has been introduced into China in recent years. The aim of this study was to evaluate the efficacy and safety of ESD in the treatment of gastrointestinal (GI) neoplasms. METHODOLOGY: Early GI neoplasms (n=41) in 40 patients from local Eastern China were treated with ESD at Zhejiang Provincial People's Hospital and followed-up from January 2009 to December 2011. Postoperative pathology, complications and therapeutic outcomes were retrospectively analyzed. RESULTS: Mean size of the resected lesions was 2.2±0.81cm (1.2-6.0cm) and mean operation time was 77±28 minutes (20-150 minutes). The rates for successful resection, en bloc resection and complications were 90.2% (37/41), 83.8% (31/37) and 9.8% (4/41), respectively. The postoperative pathology showed 4 cases of early esophageal cancer, 6 of early gastric cancer or high-grade intraepithelial neoplastic changes, 5 of rectal laterally spreading tumor, 5 of esophageal or gastric leiomyoma, 2 of gastric heterotopic pancreas, and 18 of esophageal and gastric flat lesions with low-grade intraepithelial neoplastic changes. Tumor residue or recurrence was not been detected in all 40 patients during follow-up. CONCLUSIONS: According to our experience in local Eastern China, ESD is a feasible technique for the treatment of GI neoplasms. Even though it has promising resection rate and acceptable complication rate, the indication of ESD should be selected strictly and the operators need to be well-trained.

Narrow-band imaging endoscopy with and without magnification in diagnosis of colorectal neoplasia.

AIM: To evaluate the diagnostic efficacies of narrow-band imaging (NBI) endoscopy with and without high magnification in distinguishing neoplasia from non-neoplasia colorectal lesions. METHODS: A total of 118 patients with 123 colorectal lesions examined by NBI endoscopy in the Zhejiang Provincial People's Hospital from September 2008 to April 2010 were enrolled in this study. These lesions were classified by pit pattern and capillary pattern, and then assessed by histopathology. RESULTS: Ten lesions not meeting the diagnostic criteria were excluded, the overall diagnostic accuracy of NBI endoscopy in distinguishing neoplasia from non-neoplasia colorectal lesions was 91.2% (103/113), and that of NBI endoscopy with and without high magnification was 93.0% (40/43) and 90.0% (63/70), respectively. Both were significantly higher than that of conventional colonoscopy reported in the literature (P<0.05), but there was no significant difference between the two groups (P>0.05). CONCLUSION: Besides NBI magnifying endoscopy, NBI endoscopy without magnification may also be used to distinguish neoplasia from non-neoplasia colorectal lesions.

Association of MIF-173 gene polymorphism with inflammatory bowel disease in Chinese Han population.

AIM: To study whether macrophage migration inhibitory factor (MIF)-173 gene polymorphism correlates with inflammatory bowel disease (IBD) in Chinese Han population. METHODS: MIF-173 single nucleotide polymorphism (SNP) was genotyped by tetra-primer amplification refractory mutation system (ARMS) and restriction fragment length polymorphisms (RFLP)-PCR in 142 healthy subjects and 98 patients with inflammatory bowel disease (IBD). RESULTS: There were no discrepancies between the results obtained by tetra-primer ARMS and RFLP-PCR. The frequency of MIF-173 CC genotype was significantly higher in patients with ulcerative colitis (UC) 15.5% than in healthy individuals 5.6% (chi(2)=6.066, P=0.018, OR=3.067 and 95% CI=1.257-7.482). There was a trend towards a higher frequency of CC genotype among CD patients compared with healthy controls, however this did not attain the statistical significance (P=0.245). CONCLUSION: MIF-173 CC genotype may be associated with susceptibility to UC.

[Distribution of MIF -173 single nucleotide polymorphism checked by tetra-primer amplification refractory mutation system].

OBJECTIVE: To study the distribution of macrophagemigration inhibitory factor gene (MIF) -173 single nucleotide polymorphism (SNP) of Chinese Han population in Zhejiang province. METHODS: The DNA samples were extracted from EDTA blood of 142 unrelated healthy individuals. Alleles of MIF -173 SNP were genotyped by using the techniques of tetra-primer amplification refractory mutation system (ARMS) and restriction fragment length polymorphisms (RFLP)-PCR Meantime the PCR products were cloned and sequenced. RESULTS: The authors detected three kinds of genotypes at the MIF -173 locus, and no deviation was observed from Hardy-Weinberg equilibrium. The final results were the same completely, whatever either tetra-primer ARMS or RFLP-PCR was used to check the MIF -173 single nucleotide polymorphism. Statistical analysis showed that the distributions of MIF -173 SNP alleles and genotype frequencies were significantly different between Chinese Han population and European Caucasian(P< 0.01), but no significant difference demonstrated to happen between Chinese and Japanese(P> 0.05). CONCLUSION: Tetra-primer ARMS is an accurate, rapid and economical method for SNP genotyping.There exists ethnic difference in the distribution of MIF -173 SNP alleles.

Association of tumor necrosis factor genetic polymorphism with chronic atrophic gastritis and gastric adenocarcinoma in Chinese Han population.

AIM: To investigate the association of TNF polymorphisms with chronic atrophic gastritis (CAG) and gastric adenocarcinoma in Chinese Han patients. METHODS: The TNFa-e 5 microsatellites and 3 RFLP sites were typed using PCR technique, followed by high-voltage denaturing PAGE with silver staining and restriction enzyme digestion respectively in specimens from 53 patients with CAG and 56 patients with gastric adenocarcinoma and 164 healthy controls. The PCR products were cloned and sequenced. RESULTS: The frequency of TNF-beta Ncol*1/2 genotype was higher in patients with chronic atrophic gastritis than in healthy controls, but no significant difference was observed (60.38% vs 46.34%, P=0.076). The frequency of TNa10 allele was significantly higher in patients with chronic atrophic gastritis than in healthy controls (19.81% vs 11.89%, P=0.04). However, it did not relate to age, gender, atrophic degree or intestinal metaplasia in patients with chronic atrophic gastritis. The frequency of TNF-beta Ncol*1/2 and d2/d6 genotypes were significantly higher in patients with gastric adenocarcinoma than in healthy individuals (P>0.05). However, TNF-beta Ncol*1/2 and d2/d6 genotypes did not relate to age, gender, grade of differentiation and clinicopathologic stage in patients with gastric adenocarcinoma. The frequency of TNFa6b5c1 haplotype homozygote was significantly lower in patients with gastric adenocarcinoma than in healthy controls (1.79% vs 15.85%, P=0.006). CONCLUSION: TNFa10 allele may be a risk factor for chronic atrophic gastritis. TNF-beta Ncol*1/2 and d2/d6 genotypes are associated with the susceptibility to gastric adenocarcinoma, whereas TNFa6b5c1 haplotype homozygote may contribute to the resistance against gastric adenocarcinoma.

Polymorphisms at the TNF locus in Chinese Han population.

One hundred sixty-four unrelated healthy individuals from Chinese Han population were investigated in order to define the distribution of eight polymorphic loci within the tumor necrosis factor (TNF) gene cluster and determine their relationship between the high polymorphic microsatellite TNFa, b, d, and other elements. The cloning and sequencing for five microsatellites were simultaneously done. In this study, the distribution of TNF alleles apparently vary from other ethnic groups. A new allele was detected and confirmed. It should be emphasized that a very strong association between TNFd8 and TNFe4 is reported and d8e4 haplotype appears to be specific to the population studied. In addition, five extended haplotypes were established in this population: a6b5c1d8e4TNF308-1TNF-betaNco1-1TNFAspH1-2, a2b1c2d5e1TNF308-1TNF-betaNco1-2TNFAspH1-2, a11b4c1d4e3TNF308-1TNF-betaNco1-2TNFAspH1-1, a10b4c1d4e3TNF308-1TNF-betaNco1-2TNFAspH1-1, and a2b3c1d2e3TNF308-2TNFAspH1-2. Data suggest that important ethnic differences may exist and that it is a necessary initiative for further research.