Activation of ASIC3/ERK pathway by paeoniflorin improves intestinal fluid metabolism and visceral sensitivity in slow transit constipated rats.

Slow transit constipation (STC) is one of the most common gastrointestinal disorders in children and adults worldwide. Paeoniflorin (PF), a monoterpene glycoside compound extracted from the dried root of Paeonia lactiflora, has been found to alleviate STC, but the mechanisms of its effect remain unclear. The present study aimed to investigate the effects and mechanisms of PF on intestinal fluid metabolism and visceral sensitization in rats with compound diphenoxylate-induced STC. Based on the evaluation of the laxative effect, the abdominal withdrawal reflex test, enzyme-linked immunosorbent assay, quantitative real-time polymerase chain reaction, western blot, and immunohistochemistry were used to detect the visceral sensitivity, fluid metabolism-related proteins, and acid-sensitive ion channel 3/extracellular signal-regulated kinase (ASIC3/ERK) pathway-related molecules. PF treatment not only attenuated compound diphenoxylate-induced constipation symptoms and colonic pathological damage in rats but also ameliorated colonic fluid metabolic disorders and visceral sensitization abnormalities, as manifested by increased colonic goblet cell counts and mucin2 protein expression, decreased aquaporin3 protein expression, improved abdominal withdrawal reflex scores, reduced visceral pain threshold, upregulated serum 5-hydroxytryptamine, and downregulated vasoactive intestinal peptide levels. Furthermore, PF activated the colonic ASIC3/ERK pathway in STC rats, and ASIC3 inhibition partially counteracted PF's modulatory effects on intestinal fluid and visceral sensation. In conclusion, PF alleviated impaired intestinal fluid metabolism and abnormal visceral sensitization in STC rats and thus relieved their symptoms through activation of the ASIC3/ERK pathway.

Comprehensive characterization of TGFB1 across hematological malignancies.

TGFB1, which encodes TGF-ß1, a potent cytokine regulating varies cellular processes including immune responses. TGF-ß1 plays context-dependent roles in cancers and is increasingly recognized as a therapeutic target to enhance immunotherapy responses. We comprehensively evaluated expression of TGFB1 and its clinical and biological effects across hematological malignancies. TGFB1 expression was first explored using data from the GTEx, CCLE, and TCGA databases. The expression and clinical significances of TGFB1 in hematological malignancies were analyzed using Hemap and our In Silico curated datasets. We also analyzed the relationship between TGFB1 with immune scores and immune cell infiltrations in Hemap. We further assessed the value of TGFB1 in predicting immunotherapy response using TIDE and real-world immunotherapy datasets. TGFB1 showed a hematologic-tissue-specific expression pattern both across normal tissues and cancer types. TGFB1 expression were broadly dysregulated in blood cancers and generally associated with adverse prognosis. TGFB1 expression were associated with distinct TME properties among different blood cancer types. In addition, TGFB1 expression was found to be a useful marker in predicting immunotherapy responses. Our results suggest that TGFB1 is broadly dysregulated in hematological malignancies. TGFB1 might regulate the immune microenvironment in a cancer-type-specific manner, which could be applied in the development of new targeted drugs for immunotherapy.

ceRNA regulatory network of FIH inhibitor as a radioprotector for gastrointestinal toxicity by activating the HIF-1 pathway.

Given the relentless renewal ability of intestinal crypt-base stem cells, small intestine in the gastrointestinal (GI) tract is more vulnerable to radiation-induced disruption. Through promoting epithelial integrity and reducing intracellular reactive oxygen species (ROS) levels, hypoxia-inducible factors (HIFs) have been proved to exhibit radioprotective effects in the GI tract. Therefore, enhancing stability or transcriptional activity of HIFs might be a therapeutic strategy for developing radioprotectors. Factor inhibiting HIF (FIH or HIF-1AN) can hamper transcriptional capacity of HIF-1α via interacting with Asn803 in its C-terminal domain. Previously, we discovered promoting HIF-1α transcriptional activity in vitro by FIH inhibitor-N-oxalyl-D-phenylalanine (NOFD) exerts radioprotection on cells. However, the radioprotective effect of FIH inhibitor on the GI tract and its competing endogenous RNA (ceRNA) regulatory network from the FIH/HIF axis has never been addressed. Here we verified radioprotection of NOFD for the GI tract by an animal model and performed whole-transcriptome analysis to fully elucidate the radioprotective mechanism from the FIH/HIF axis against GI syndrome. We identified two novel circular RNAs (circRNAs) (circRNA_2909 and circRNA_0323) and two long non-coding RNAs (lncRNAs) (NONMMUT140549.1 and NONMMUT148249.1) that promote expression of HIF1A and NOS2 in the HIF-1 pathway by sponging microRNAs (miRNAs), especially mmu-miR-92a-1-5p. The de-repression of HIF-1α transcriptional capacity by inhibiting FIH proteomic activity suggests a new therapeutic strategy in alleviating radiation-induced GI syndrome.

Comparisons of perioperative and survival outcomes of laparoscopic versus open gastrectomy for serosa-positive (pT4a) gastric cancer patients: a propensity score matched analysis.

BACKGROUND: Data about whether laparoscopic gastrectomy (LG) is applicable in serosa-positive (pT4a) gastric cancer patients remain rare. The purpose of this study is to compare the perioperative and long-term outcomes between the laparoscopic and open gastrectomy (OG) in pT4a gastric cancer patients who underwent curative resection. METHODS: A total of 1086 consecutive pT4a patients (101 patients with LG and 985 with OG) who underwent curative gastrectomy in a high-volume center between 2006 and 2016 were evaluated. Demographics, surgical, and oncologic outcomes were analyzed. Propensity score matching (PSM) analysis was performed to balance baseline confounders, and COX regression analysis was performed to identify independent prognostic factors. RESULTS: After PSM adjustment, a well-balanced cohort comprising 101 patients who underwent LG and 201 who underwent OG was analyzed. Operative time (288.7 vs. 234.2 min; P < 0.001) was significantly longer, while estimated blood loss (172.8 vs. 220.7 ml; P < 0.001) was significantly less in the LG group compared with the OG group. There were no significant differences between groups in total number of harvested lymph nodes, postoperative stays, readmission rate, and postoperative complication rate. The 3-year overall survival (OS) rate was not significant different in the LG and OG groups (66.7% vs. 62.8%, P = 0.668), and the subsequent multivariate analysis revealed that the surgical approach was not an independent prognostic factor for OS (HR = 1.123; 95%CI: 0.803-1.570; P = 0.499). In sensitivity analysis including 78 pairs well-matched patients operated by an experienced surgeon, the results were similar to these for the matched entire cohort. CONCLUSION: LG can be a safe and feasible approach for pT4a gastric cancer treatment. However, well-designed high-quality RCTs are expected to draw a definitive conclusion on this topic.

Copper-Catalyzed Trifluoromethylation/Cyclization of Alkynes for Synthesis of Dioxodibenzothiazepines.

A facile and efficient approach for the synthesis of the CF3-containing dioxodibenzothiazepines has been developed via copper-catalyzed trifluoromethylation/cyclization of alkynes utilizing a radical relay strategy. This method has demonstrated low catalyst loading, high regiocontrol, and broad scope under mild conditions. Good compatibility for the N-protecting group, gram-scale experiment, and further derivation of product prove the versatility of this transformation.

Enantioselective Copper-Catalyzed 1,5-Cyanotrifluoromethylation of Vinylcyclopropanes.

A copper-catalyzed enantioselective 1,5-cyanotrifluoromethylation of vinylcyclopropanes has been developed using a radical relay strategy. This asymmetric reaction has demonstrated high enantioselective control, broad substrate scope, and mild conditions. Initiated by the in situ generated CF3 radical from Togni's reagent, this method offers a new solution for remote enantioselective bifunctionalization of alkenes and thus provides a straightforward way for the synthesis of chiral CF3-containing internal alkenylnitriles.

Nickel-catalyzed monofluoromethylation of (hetero)aryl bromides via reductive cross-coupling.

A mild and efficient nickel-catalyzed direct monofluoromethylation of (hetero)aryl bromides by reductive cross-coupling has been developed. This method exhibits good efficiency, wide functional-group compatibility, and suitability for aryl and heteroaryl bromides with abundant industrial raw material BrCH2F. This strategy provides an efficient way to synthesize monofluoromethylated molecules for drug discovery.

Mesenchymal Stem Cells Induced by Microencapsulated Chondrocytes on Repairing of Intervertebral Disc Degeneration.

OBJECTIVE: To evaluate the therapeutic effects of mesenchymal stem cells induced by microencapsulated chondrocytes on repairing of intervertebral disc degeneration. METHODS: Rabbit chondrocytes and bone marrow-derived mesenchymal stem cells (MSC) were derived. Chondrocytes were microencapsulated by a microcapsule generator to produce microencapsulated chondrocytes (MEC). MSC were then co-cultured with MEC (MSC-MEC) and the properties and the therapeutic effects on repairing of intervertebral disc degeneration were studied. For the in vitro study, cell proliferation, type II collagen, and glycosaminoglycan (GAG) were studied. The MSC induced by chondrocytes in the Transwell system (MSC-MLC) and pure MSC were used as the control group. For the in vivo studied, MSC-MEC were implanted into the intervertebral disc degenerated (IDD) models, and the radiological images, biomechanical properties, collagen II, and histology of the discs were studied. The IDD, MSC, and MSC-MLC groups were used as the control group. RESULTS: In the in vitro study, no significant differences were found among the three groups, indicating that the microcapsule co-culture system will not affect the proliferation of MSC. The type II collagen quantity secreted by MSC-MEC was 23.57 ± 2.46 ng/µL, which was more than for MSC-MLC (15.14 ± 2.31 ng/µL) and MSC groups (4.17 ± 1.23 ng/µL, all P < 0.025). GAG secreted by MSC-MEC was 0.184 ± 0.006 mg/well, which was more than for the MSC-MLC (0.151 ± 0.011 mg/well) and MSC groups (0.023 ± 0.002 mg/well, all P < 0.025). In the in vivo study, no obvious degenerative or protrusive disc was found in the MSC-MEC group, while protrusive discs could be found in the MSC-MLC group, and both degenerative and protrusive discs were found in MSC and IDD groups, which indicated that the reparative effects of MSC-MEC on degenerated discs were better than for the control groups. Biomechanical properties of discs in the MSC-MEC group were maintained at all four time points (2nd, 4th, 8th, and 16th week after implantation). The compressive strength (CS) and the elastic modulus (EM) of MSC and IDD groups were consistently decreased. The CS of the MSC-MLC group was increased in the 4th week but decreased again in the 8th week, while the EM of the MSC-MLC group consistently decreased. Western blot results indicated that discs of the MSC-MEC group had more collagen II, which is an important component of discs. Histology staining showed that the nucleus pulposus of MSC-MEC was complete; no obvious fragment of component loss was found, while those of MSC-MLC, MSC, and IDD groups were widened, broken, and hollow. CONCLUSION: The microencapsulation method for half-contact co-culturing improves the differentiation extent of MSC, and MSC induced by chondrocytes could also be used for treatment of IDD.

Recombinant expression, purification and antimicrobial activity of a novel antimicrobial peptide PaDef in Pichia pastoris.

The antimicrobial peptide PaDef was isolated from Mexican avocado fruit and was reported to inhibit the growth of Escherichia coli and Staphylococcus aureus in 2013. In this study, an N-terminal 6 × His tagged recombinant PaDef (rPaDef) with a molecular weight of 7.5 KDa, for the first time, was expressed as a secreted peptide in Pichia pastoris. The optimal culture condition for rPaDef expression was determined to be incubation with 1.5% methanol for 72 h at 28 °C under pH 6.0. Under this condition, the amount of the rPaDef accumulation reached as high as 79.6 µg per 1 ml of culture medium. Once the rPaDef peptide was purified to reach a 95.7% purity using one-step nickel affinity chromatography, its strong and concentration-dependent antimicrobial activity was detected to be against a broad-spectrum of bacteria of both Gram-negative and Gram-positive. The growth of these bacterial pathogens was almost completely inhibited when the rPaDef peptide was at a concentration of as low as 90 µg/ml. In summary, our data showed that rPaDef derived from Mexican avocado fruit can be expressed and secreted efficiently when P. pastoris was used as a cell factory. This is the first report on heterologous expression of PaDef in P. pastoris and the approach described holds great promise for antibacterial drug development.

Expression, purification and initial characterization of a novel recombinant antimicrobial peptide Mytichitin-A in Pichia pastoris.

Mytichitin-A is an antimicrobial peptide isolated from the serum of Mytilus coruscus and is reported to inhibit bacterial growth as tested on several Gram-positive bacteria. To produce large quantity of Mytichitin-A to further investigate its biological activity, nucleotide sequence encoding a recombinant 6 × His-Mytichitin-A (rMytichitin-A) peptide was synthesized and inserted into the inducible yeast expression vector pPICZαA. With the availability of such an expression vector called pPICZαA-Mytichitin-A, we transformed Pichia pastoris GS115 cells with a SacI-linearized pPICZαA-Mytichitin-A by electroporation. Transgenic strains secreting rMytichitin-A with a molecular weight of approximate 10 KDa as expected were obtained. The optimal culture condition for rMytichitin-A expression was determined to be 1.0% methanol induction, 96 h incubation at 28 °C and the amount of rMytichitin-A reached 45.5 µg/ml. The percentage of rMytichitin-A was estimated to be 73.6% of the total protein. After rMytichitin-A was purified using nickel ions affinity chromatography, approximate 9.1 mg pure rMytichitin-A was obtained from 500 ml of cell culture medium with 97.8% purity. More importantly, both the culture supernatant and purified rMytichitin-A inhibited the growth of Gram-positive bacteria, especially Staphylococcus aureus and Bacillus subtilis with a minimum inhibition concentration of as low as 31 and 48 µg/ml, respectively. Differently from the native protein, however, the rMytichitin-A is not active against Gram-negative bacteria. Taken together, this is the first report on the heterologous expression of Mytichitin-A in P. pastoris. Our study showed that P. pastoris is an effective expression system for producing large quantities of biologically active Mytichitin-A for both research and application purposes.

[Proximal femoral geometry changes after femoral neck fracture treated with total hip arthroplasty].

OBJECTIVE: To observe the changes of proximal femoral geometry after femoral neck fracture treated with THA, analyze the existent of differences and their manifestation. METHODS: All patients of femoral neck fracture (FNF) and osteonecrosis of femoral head (ONFH) were treated with THA by the same operating team from January to December of 2014, including 22 patients with FNF (11 males and 11 females,with age from 44 to 83 years old (means 66.18 ± 11.47) and 23 patients with ONFH (12 males and 11 females, with age from 19 to 68 years old (means 51.91 ± 11.76). After THA, height of femorals, offsets, osteotomy position and adjusting modes were measured and the statistic analysis was done. RESULTS: After THA, all patients were measured. Decreased femoral height, offsets and lower osteotomy positions were found in patients with FNF than those with ONFH, and 3 kinds of adjustments because of lower-positional osteotomy were found. CONCLUSION: After THA, lower-positional osteotomy and decreased femoral offsets may occur on patients with FNF. The adjustments caused by lower-positional osteotomy may lead to negative results.

Evaluation of colonoscopy in the diagnosis and treatment of rectal carcinoid tumors with diameter less than 1 cm in 21 patients.

The aim of this study was to evaluate colonoscopy in the diagnosis and treatment of rectal carcinoid tumors with diameter <1 cm. Elevated lesions with normal mucosal appearance under colonoscopy were identified. Endoscopic ultrasound (EUS) was performed in 16 patients. Lesions diagnosed as rectal carcinoid tumors were resected by endoscopic mucosal resection (EMR). The diagnosis of specimens by EMR was confirmed by pathological examination. Immunohistochemical staining was undertaken and follow-up data were collected. Twenty-two lesions were found among the 21 cases. The majority of these were located within 10 cm of the anal opening. Twenty two cases with rectal carcinoids were diagnosed by EUS under colonoscopy and all cases were verified by pathological examination. The resection rate was 95.5% (21/22). Of the lesions, six were mucosal and 10 were submucosal. Immunohistochemistry was undertaken for carcinoid tumors. Histological patterns of rectal carcinoids revealed solid nests or trabecular patterns. Eleven cases were synaptophysin (SYN)-positive, 8 cases were neurone-specific enolase (NSE)-positive and 5 cases were chromogranin A (CgA)-positive. Colonoscopy combined with EUS is effective in the diagnosis and determination of small rectal carcinoids. Endoscopic treatment is effective for small-sized tumors. Pathology and immunohistochemistry remain the diagnostic gold standard.

Moxifloxacin-containing triple therapy versus bismuth-containing quadruple therapy for second-line treatment of Helicobacter pylori infection: a meta-analysis.

BACKGROUND: Moxifloxacin-containing triple therapy has been suggested as an alternative second-line therapy for Helicobacter pylori infection. AIMS: To systematically review the efficacy and tolerance of moxifloxacin-containing triple therapy in second-line H. pylori eradication, and to conduct a meta-analysis of studies comparing this regimen with bismuth-containing quadruple therapy. MATERIALS AND METHODS: Electronic databases including Medline, Embase, Cochrane controlled trials register, Web of Science, PubMed, Chinese Biomedical Literature Database (updated to December 2010), and manual searches were conducted. A meta-analysis of all randomized controlled trials (RCTs) comparing moxifloxacin-containing triple therapy to bismuth-containing quadruple therapy in the second-line treatment of H. pylori infection was performed. RESULTS: Seven RCTs including 787 patients were assessed. The meta-analysis showed that the eradication rate in the moxifloxacin group was significantly higher than that in the quadruple therapy group (74.9 vs 61.4%, OR 1.89, 95% CI: 1.38-2.58, p < .0001); besides, the rates of side effects and discontinuing therapy because of side effects in the moxifloxacin group were significantly lower than those in the quadruple therapy group (side effects: 10.1 vs 27.8%, OR 0.27, 95% CI: 0.18-0.41, p < .00001; discontinuing therapy because of side effects: 1.4 vs 8.2%, OR 0.18, 95% CI: 0.08-0.40, p < .0001). These results were constant in the sensitivity analyses. CONCLUSION: Moxifloxacin-containing triple regimen is more effective and better tolerated than the bismuth-containing quadruple therapy in the second-line treatment of H. pylori infection.

Detection of K-ras gene mutation in fecal samples from elderly large intestinal cancer patients and its diagnostic significance.

AIM: To study the diagnostic significance of K-ras gene mutations in fecal samples from elderly patients with large intestinal cancer. METHODS: DNA was extracted in the fecal and tissue samples from 23 large intestinal cancer patients, 20 colonic adenomatoid polypus patients and 20 healthy subjects. The K-ras gene mutations at the first and second bases of codon 12 were detected by the allele specific mismatch method. RESULTS: The K-ras gene mutation was 56.52%(13/23) in the large intestinal cancer patients, which was notably higher than that in the normal subjects whose K-ras gene mutation was 5%(1/20) (chi (2)=12.93, P<0.001). There was no significant difference in comparison with that of colonic adenomatoid polypus patients whose K-ras gene mutation was 30%(6/12) (chi (2)=3.05, P>0.05). The K-ras gene mutation at the second base of codon 12 was 92.13%(12/13) in the large intestinal cancer patients. There was no significant difference between the detection rate of K-ras gene mutation in the fecal and tissue samples (chi (2)=9.35, P<0.01). CONCLUSION: Our results indicate that detection of the K-ras gene mutations in fecal samples provides a non-invasive diagnostic method for the elderly large intestinal cancer patients. Its significance in the early diagnosis of large intestinal cancer awaits further studies.

Colonoscopic screening and follow-up for colorectal cancer in the elderly.

AIM: To improve the prevention and treatment of senile patients with colorectal cancer by evaluating the importance of colonoscopy in clinical screening and follow-up. METHODS: Clinical screening of colonoscopy was performed for 2196 patients aged 60-90 years old according to the protocol,and 1740 of them (79.2%) were followed-up. RESULTS: Colorectal cancer was found in 52 patients, and the detectable rate was 2.4%. Among them, 19 were diagnosed as early colorectal cancer, accounting for 36.5% of the detected colorectal cancer. Among the followed-up patients, early colorectal cancer was found in 9, accounting for 45.0% of the detected colorectal cancer. The resectable rate and 5 years survival rate of colorectal cancer were 97.7% and 80.9% respectively. The incidence of complication was 0.05%, and the successful rate of cecum intubation was 98.9%. CONCLUSION: Colonoscopic screening and follow-up of the elderly for colorectal cancer and pre-cancerous lesion (adenomatoid polyp) can increase the detectable rate of early colorectal cancer and improve its prevention and treatment.