Chinese physicians' perceptions of fecal microbiota transplantation

AIM: To explore Chinese physicians' perceptions towards fecal microbiota transplantation(FMT) and to provide information and an assessment of FMT development in China.METHODS: A self-administered questionnaire was developed according to the FMT practice guidelines and was distributed to physicians in hospitals via Internet Research Electronic Data Capture(REDcap) software and electronic mails to assess their attitudes toward and knowledge of FMT. The questionnaire included a brief introduction of FMT that was followed by 20 questions. The participants were required to respond voluntarily, under the condition of anonymity and without compensation. Except for the fill-in-the-blank questions, all of the other questions were required in the REDcap data collection systems, and the emailed questionnaires were completed based on eligibility.RESULTS: Up to December 9, 2014, 844 eligible questionnaires were received out of the 980 distributed questionnaires, with a response rate of 86.1%. Among the participants, 87.3% were from tertiary hospitals, and there were 647(76.7%) gastroenterologists and 197(23.3%) physicians in other departments(nongastroenterologists). Gastroenterologists' awareness of FMT prior to the survey was much higher than non-gastroenterologists'(54.3 vs 16.5%, P < 0.001); however, acceptance of FMT was not statistically different(92.4 vs 87.1%, P = 0.1603). Major concerns of FMT included the following: acceptability to patients(79.2%), absence of guidelines(56.9%), and administration and ethics(46.5%). On the basis of understanding, the FMT indications preferred byphysicians were recurrent Clostridium difficile infection(86.7%), inflammatory bowel disease combined with Clostridium difficile infection(78.6%), refractory ulcerative colitis(70.9%), ulcerative colitis(65.4%), Crohn's disease(59.4%), chronic constipation(43.7%), irritable bowel syndrome(39.1%), obesity(28.1%) and type 2 diabetes(23.9%). For donor selection, the majority of physicians preferred individuals with a similar gut flora environment to the recipients. 76.6% of physicians chose lower gastrointestinal tract as the administration approach. 69.2% of physicians considered FMT a safe treatment. CONCLUSION: Chinese physicians have awareness and a high acceptance of FMT, especially gastroenterologists, which provides the grounds and conditions for the development of this novel treatment in China. Physicians' greatest concerns were patient acceptability and absence of guidelines.

Gemcitabine and cisplatin combined with the Chinese herbal medicine compound Fuzhenggubenfang improve quality of life and progression-free survival in patients with advanced non-small cell lung cancer

Objective： To assess the role of chemotherapy combined with the compound Chinese herbal medicine,Fuzhenggubenfang （FZGBF）, for treating advanced non-small-cell lung cancer. Methods： A total of 84 eligible patientswere enrolled from October 2013 to July 2016. Patients were randomized to receive either chemotherapy alone as thecontrol group or chemotherapy combined with FZGBF as the experimental group. The primary endpoint of the study wasquality of life （QOL） and progression-free survival （PFS）. Secondary endpoints were tumor response rate, toxicity,dropout rate, and univariate and multivariate analyses of clinicopathologic factors for QOL and PFS. Results： There wasa significant improvement in QOL, including better overall health （P 〈 0.001）, physical function （P 〈 0.001）, rolefunction （P 〈 0.001）, emotional function （P 〈 0.001）, cognitive function （P 〈 0.001）, and social function （P = 0.031）.Less fatigue, nausea or vomiting, insomnia, appetite loss, constipation, and alopecia were noted （All P 〈 0.001） whenFZGBF was combined with chemotherapy in comparison to chemotherapy alone. The experimental group had a betterPFS compared with the control group （P = 0.032）. There was no significant difference in tumor response rate. FZGBFsignificantly reduced chemotherapy-induced anemia （P 〈 0.001）, neutropenia （P = 0.023）, nausea and vomiting （P 〈0.001）. The use of Chinese herbal compounds had only mild side effects. In this study, factors influencing QOL were theuse of the Chinese herbal compounds （P 〈 0.001）, performance status score （P = 0.027）, clinical staging of cancer （P =0.009）, and sex （P = 0.044）. Use of traditional Chinese medicine （P = 0.043） and the number of previous chemotherapysessions （P = 0.003） were the factors influencing PFS in this study. Conclusion： FZGBF could improve QOL,compliance to treatment, relieved chemotherapy-related toxicities of patients, and consequently improved PFS, which isa promising drug combination in complementary medicine for the treatment of advanced NSCLC.

Protein expression of sensory and motor nerves Two-dimensional gel electrophoresis and mass spectrometry

The present study utilized samples from bilateral motor branches of the femoral nerve, as well as saphenous nerves, ventral roots, and dorsal roots of the spinal cord, to detect differential protein expression using two-dimensional gel electrophoresis and nano ultra-high performance liquid chromatography electrospray ionization mass spectrometry tandem mass spectrometry techniques. A mass spectrum was identified using the Mascot search. Results revealed differential expression of 11 proteins, including transgelin, Ig kappa chain precursor, plasma glutathione peroxidase precursor, an unnamed protein product （gil55628）, gfyceraldehyde-3-phosphate dehydrogenase-like protein, lactoylgfutathione lyase, adenyfate kinase isozyme 1, two unnamed proteins products （gil55628 and gi11334163）, and poly（rC）-binding protein 1 in motor and sensory nerves. Results suggested that these proteins played roles in specific nerve regeneration following peripheral nerve injury and served as specific markers for motor and sensory nerves.

BMP-6 inhibits microRNA-21 expression in breast cancer through repressing 6EF1 and AP-1

MicroRNAs （miRNAs）, which are small noncoding RNA molecules, play important roles in the post-transcriptional regulation process. The microRNA-21 gene （miR-21） has been reported to be highly expressed in various solid tumors, including breast cancer. Bone morphogenetic protein-6 （BMP-6） has been identified as an inhibitor of breast cancer epithelial-mesenchymal transition （EMT） through rescuing E-cadherin expression. We initiated experi- ments to identify the relationships between miR-21 and BMP-6 in breast cancer progression. Real-time PCR analysis showed that miR-21 expression was very high in MDA-MB-231 cells that expressed little BMP-6. A reverse correla- tion between BMP-6 and miR-21 was also determined in breast cancer tissue samples. Moreover, BMP-6 inhibited miR-21 transcription in MDA-MB-231 cells. In order to investigate how BMP-6 inhibited the miR-21 promoter （miPPR-21）, we constructed a series of miPPR-21 reporters. Luciferase assay results indicated that BMP-6 inhibited miPPR-21 activity through the E2-box and AP-l-binding sites. We also demonstrated that both δEF1 and TPA in- duced miR-21 expression. Using site-directed mutation and CHIP assay, we found that δEF1 induced miPPR-21 ac- tivity by binding to the E2-box on miPPR-21. Moreover, TPA triggered miPPR-21 activity through the AP-I binding sites. BMP-6 treatment significantly reduced the binding of these factors to miPPR-21 by decreasing the expression of δEF1 and c-Fos/c-Jun. We also demonstrated that BMP-6-induced downregulation of miR-21 modified the activ- ity of PDCD4 3＇UTR and inhibited MDA-MB-231 cell invasion. δEF1 overexpression and TPA induction blocked this inhibitory effect of BMP-6. In conclusion, BMP-6-induced inhibition of miR-21 suggests that BMP-6 may function as an anti-metastasis factor by a mechanism involving transcriptional repression of miR-21 in breast cancer.

Epigenetic changes in colorectal cancer

Epigenetic changes frequently occur in human colorectal cancer.Genomic global hypomethylation,gene promoter region hypermethylation,histone modifications,and alteration of miRNA patterns are major epigenetic changes in colorectal cancer.Loss of imprinting(LOI) is associated with colorectal neoplasia.Folate deficiency may cause colorectal carcinogenesis by inducing gene-specific hypermethylation and genomic global hypomethylation.HDAC inhibitors and demethylating agents have been approved by the FDA for myelodysplastic syndrome and leukemia treatment.Non-coding RNA is regarded as another kind of epigenetic marker in colorectal cancer.This review is mainly focused on DNA methylation,histone modification,and microRNA changes in colorectal cancer.

Wnt5a Signaling A New and Attractive Target for Specific Anticancer Therapy

Wnt signaling has been shown to engage a multifunctional pathway that is involved in the regulation of a wide variety of normal and pathologic processes, including embryogenesis, differentiation and tumorigenesis. Recent studies have demonstrated that Wnt5a expression is frequently seen in various human cancers. In contrast to the transforming members of the Wnt family, shown to be upregulated in many cancers, the role of Wnt5a is still controversial in its expression in different tumors. There is increasing evidence that Wnt5a has tumor suppressor function in some malignancies, and in addition, it elicits promigratory and proinvasive effects via the planar cell polarity pathway, which suggests that Wnt5a might be an effective marker for the progression and prognosis of tumors. Obviously, the outcome of Wnt5a signaling is dependent on a multitude of variables, ranging from receptors, downstream effectors and inhibitors, to external influences coming from the tumor microenvironment. This review will focus on the role of Wnt5a signaling and, as a consequence, provide an outline describing the expression and functions of Wnt5a in cancer progression.

Mesenchymal stem cell-derived exosomes inhibit the VEGF-A expression in human retinal vascular endothelial cells induced by high glucose

AIM:To determine the effect of exosomes derived from human umbilical cord blood mesenchymal stem cells(hUCMSCs)on the expression of vascular endothelial growth factor A(VEGF-A)in human retinal vascular endothelial cells(HRECs).METHODS:Exosomes were isolated from hUCMSCs using cryogenic ultracentrifugation and characterized by transmission electron microscopy,Western blotting and nanoparticle tracking analysis.HRECs were randomly divided into a normal control group(group A),a high glucose model group(group B),a high glucose group with 25μg/mL(group C),50μg/mL(group D),and 100μg/mL exosomes(group E).Twenty-four hours after coculture,the cell proliferation rate was detected using flow cytometry,and the VEGF-A level was detected using immunofluorescence.After coculture 8,16,and 24h,the expression levels of VEGF-A in each group were detected using PCR and Western blots.RESULTS:The characteristic morphology(membrane structured vesicles)and size(diameter between 50 and 200 nm)were observed under transmission electron microscopy.The average diameter of 122.7 nm was discovered by nanoparticle tracking analysis(NTA).The exosomal markers CD9,CD63,and HSP70 were strongly detected.The proliferation rate of the cells in group B increased after 24h of coculture.Immunofluorescence analyses revealed that the upregulation of VEGF-A expression in HRECs stimulated by high glucose could be downregulated by cocultured hUCMSC-derived exosomes(F=39.03,P<0.01).The upregulation of VEGF-A protein(group C:F=7.96;group D:F=17.29;group E:F=11.89;8h:F=9.45;16h:F=12.86;24h:F=42.28,P<0.05)and mRNA(group C:F=4.137;group D:F=13.64;group E:F=22.19;8h:F=7.253;16h:F=16.98;24h:F=22.62,P<0.05)in HRECs stimulated by high glucose was downregulated by cocultured hUCMSC-derived exosomes(P<0.05).CONCLUSION:hUCMSC-derived exosomes downregulate VEGF-A expression in HRECs stimulated by high glucose in time and concentration dependent manner.

Hypoxia inducible factor 1α promotes interleukin-1 receptor antagonist expression during hepatic ischemia-reperfusion injury

BACKGROUND Ischemia-reperfusion injury(IRI) is a major risk associated with liver surgery and transplantation,and its pathological mechanism is complex.Interleukin-1 receptor antagonist(IL-1ra) can protect the liver from IRI.However,the regulatory mechanism of IL-1ra expression is still unclear.AIM To identify the mechanism that could protect the liver in the early stage of IRI.METHODS To screen the key genes in hepatic IRI,we performed RNA sequencing and gene enrichment analysis on liver tissue from mice with hepatic IRI.Subsequently,we verified the expression and effect of IL-1ra in hepatic IRI.We also used promoter mutagenesis and chromatin immunoprecipitation assay to search for the transcriptional regulatory sites of hypoxia-inducible factor(HIF)-1α.Finally,to explore the protective mechanism of ischemic preconditioning(IP),we examined the expression of HIF-1α and IL-1ra after IP.RESULTS We identified IL-1ra as a key regulator in hepatic IRI.The expression of IL-1ra was significantly upregulated after hepatic IRI both in vivo and in vitro.Furthermore,we found that HIF-1αregulated Il-1ra transcription in response to hypoxia.Increased HIF-1α accumulation promoted IL-1ra expression,whereas inhibition of HIF-1α exhibited the opposite effect.We also confirmed a predominant role for hypoxia response element in the regulation of Il1ra transcription by HIF-1αactivation.Of note,we demonstrated that IP protects against hepatic IRI by inducing IL-1ra expression,which is mediated through HIF-1α.CONCLUSION We demonstrated that ischemia or hypoxia leads to increased expression of IL-1ra through HIF-1α.Importantly,IP protects the liver from IRI via the HIF-1α–IL-1ra pathway.

A novel tissue engineered nerve graft constructed with autologous vein and nerve microtissue repairs a longsegment sciatic nerve defect

Veins are easy to obtain,have low immunogenicity,and induce a relatively weak inflammatory response.Therefore,veins have the potential to be used as conduits for nerve regeneration.However,because of the presence of venous valves and the great elasticity of the venous wall,the vein is not conducive to nerve regeneration.In this study,a novel tissue engineered nerve graft was constructed by combining normal dissected nerve microtissue with an autologous vein graft for repairing 10-mm peripheral nerve defects in rats.Compared with rats given the vein graft alone,rats given the tissue engineered nerve graft had an improved sciatic static index,and a higher amplitude and shorter latency of compound muscle action potentials.Furthermore,rats implanted with the microtissue graft had a higher density and thickness of myelinated nerve fibers and reduced gastrocnemius muscle atrophy compared with rats implanted with the vein alone.However,the tissue engineered nerve graft had a lower ability to repair the defect than autogenous nerve transplantation.In summary,although the tissue engineered nerve graft constructed with autologous vein and nerve microtissue is not as effective as autologous nerve transplantation for repairing long-segment sciatic nerve defects,it may nonetheless have therapeutic potential for the clinical repair of long sciatic nerve defects.This study was approved by the Experimental Animal Ethics Committee of Chinese PLA General Hospital(approval No.2016-x9-07)on September 7,2016.

Perineural Invasion in Pancreatic Cancer： Advanced Research in the Neuro-cancer Interactions

Pancreatic Cancer （PCa） is characterized by prominently local nerve alterations and perineural invasion （PNI）, which frequently affects the extrapancreatic nerve plexus, causing severe pain and retropancreatic tumor extension. It precludes curative resection, promotes local recurrence, and at the last negatively influences the prognosis of patients. Recent research on PNI in PCa has revealed the critical involvement of numerous nerve- or cancer cell-derived molecules in vitro and in vivo. However, the mechanisms contributing to alteration and invasion of intrapancreatic nerves and the spread of cancer cells along extrapancreatic nerves in pancreatic cancer patients are still poorly understood. This review focuses on perineural invasion in pancreatic cancer and provides an outline of the characteristics and molecular mechanisms of perineural invasion in pancreatic cancer.

Inhibiting effect of Astragalus polysaccharides on the functions of CD4＋CD25^highTreg cells in the tumor microenvironment of human hepatocellular carcinoma

Background Astragalus polysaccharides （APS）, the main active extract from Astragalus membranaceus （a traditional Chinese medicinal herb）, is associated with a variety of immunomodulatory activities. The purpose of the present study was to examine the effect of APS on the function of Treg cells in the tumor microenvironment of human hepatocellular carcinoma （HCC） and to identify the pharmacologic mechanism of APS responsible for the anti-chemotactic activity in CD4＋CD25^highTreq cells in tumor site of HCC.

Effects of subdiaphragmatic cardiac compression on cardiac arrest during liver transplantation

Cardiac arrest during upper abdominal surgery such as liver transplantation is a rare but very severe complication. Traditional external cardiac compression has been the mainstay of basic life support in general circumstances. Subdiaphragmatic cardiac compression （SDCC）, with no incision in the diaphragm, may be a more effective measure. This maneuver can provide more effective and timely cardiac compression via the already open abdomen in surgery and not add extra trauma. This method can provide a quicker and more effective means of circulation support for intraoperative cardiac arrest patients without adding new injuries. Five cases are reported and all the patients had return of spontaneous circulation （ROSC）. This is the first report of the SDCC method.

Significance of low serum vitamin D for infection risk, disease severity and mortality in critically ill patients

Background Hospitalized patients often have higher rate of vitamin D deficiency than healthy people. Vitamin D levels below normal are associated with hospital stay, increased incidence of adverse prognosis and increased mortality of a number of diseases. Whether there is a relationship between vitamin D levels and infection or sepsis in the critically ill is still unclear. This study will explore the relationship between vitamin D levels and risk of infection, assessment for disease severity, and predictor of mortality. Methods To evaluate the value of vitamin D in intensive care unit （ICU） cases to sepsis, severity and prognosis assessment, high performance liquid chromatography and tandem mass spectrometry were used to measure the concentrations of vitamin D in sera of critically ill patients. The serum samples were drawn within the first 24 hours of ICU admission. Results The study included 206 people, 50 healthy controls, 51 ICU control patients and 105 ICU diagnosed with sepsis. Critically ill ICU patients （ICU sepsis and ICU control group） had lower vitamin D concentration than normal people, but septic patients showed no significant reduction of vitamin D concentration when compared with critically ill patients with no positive etiological evidence. For assessment of disease severity, there were very low negative correlations between APACHE II, SAPS II and SOFA scores and vitamin D level. Additionally, patients of different 25-（OH）D levels showed no difference whether in terms of 28-day survival （χ2=1.78, P=0.776） or 90-day survival （χ2=4.12, P=0.369）. Multivariate Logistic regression demonstrated that APECHE II and SAPS II scores were independent risk factors to deaths caused by sepsis. Conclusion Clinically, serum concentration of vitamin D is not an indicator for diagnosis and assessment in critically ill patients （ClinicalTrial.gov identifier NCT01636232）.

Heliox as a driving gas to atomize inhaled drugs on acute exacerbation of chronic obstructive pulmonary disease： a prospective clinical study

Background Acute exacerbation of chronic obstructive pulmonary disease （AECOPD） is a common condition,which affects not only the quality of life of patients but also their prognosis.The purpose of this study was to explore the effects of an inhaled salbutamol sulfate solution and an inhalation suspension of the glucocorticoid budesonide that were atomized with heliox to treat patients with AECOPD.Methods Twenty-three patients with AECOPD were divided into a treatment group （He/O2=70％/30％） and a control group （N2/O2=70％/30％）.The salbutamol sulfate and budesonide were administered by inhalation twice a day for 7 days.Vital signs,arterial blood gas levels,pulmonary function and the levels of serum myostatin （sMSTN） were measured and lung vibration imaging was performed.Results We found that the PaO2 and PaCO2 values were not significantly different between the two groups at the various time points （P ＞0.05）.There were also no significant differences in any of the parameters of pulmonary function between the two groups.However,after baseline correction,the increase rate of the forced expiratory volume in one second （FEV1）,the forced vital capacity （FVC）,and the maximum minute ventilation （MW） appeared to be significantly increased at some time points compared with the baseline （before treatment） in both groups （P ＜0.05）.Although the values of quantitative lung distribution （QLD） for different regions and the levels of sMSTN were slightly different between the two groups,the repeated measures analysis of variance （ANOVA） revealed that there were no significant differences between the two groups or within any group （P ＞0.05）.Conclusion Although the use of heliox as a driving gas can improve symptoms and benefit patients with AECOPD,the heliox treatment group did not have significant differences in arterial blood gases,lung function,lung vibration response imaging or the levels of sMSTN compared with the control group.（Chinese Clinical Trial Register Center ChiCTR-TRC-00000273）

Effect of antiallergic herbal agents on chloride channel-3 and immune microenvironment in nasal mucosal epithelia of allergic rhinitis rabbits

Background Allergic rhinitis （AR） is a Th2 dominant cytokine response. Chloride channel-3 （CIC-3） plays an important role in nasal mucosal edema and inflammatory pathologic changes in AR. Antiallergic herbal agents （AHA） are antiallergic herbal products. In the previous study, we have demonstrated that AHA clearly inhibited allergic medium and relieved allergic reaction of AR. The aim of this study was to evaluate the function of CIC-3 and discuss the possible therapeutic effects of AHA on immune microenvironment in AR. Methods AHA were produced and used to treat AR. An animal model of an AR rabbit was established by ovalbumin （OVA）. The rhinitis rabbits were randomly divided into three groups： AHA treated group （AHATG）, model group （MG） and healthy control group （HCG）. The expressions of CIC-3 protein were examined by immunohistochemical method. The mucosal epithelial cells of all the rabbit groups were primarily cultured with tissue culture method in vitro with or without rhlL-4 or rhlL-2. Furthermore, the expressions of CIC-3 mRNA were detected by real-time PCR. The levels of monocyte chemotactic factor-1 （MCP-1） and vascular cell adhesion molecule-1 （VCAM-1） protein in culture supernatants were measured by ELISA. Results The expressions of CIC-3 mRNA increased more in mucosal epithelial cells of MG than those in AHATG and HCG （P 〈0.01）. The levels of CIC-3 mRNA, MCP-1 and VCAM-1 protein in culture supernatants of MG were significantly higher than those in the other two groups （P 〈0.01）. Those were significantly increased in MG untreated 12 hours later than those in other two groups （P 〈0.01）. The expressions of CIC-3 mRNA, MCP-1 and VCAM-1 protein in culture supernatants of MG and HCG treated with rhlL-4 were significantly higher than those in the AHATG treated with rhlL-4 （P 〈0.01）. The levels of CIC-3 mRNA, MCP-1 and VCAM-1 protein in culture supernatants of all groups treated with rhlL-2 showed no significant changes （P 〉0.05）. Conclusions AHA can inhibit the secretions of CIC-3, MCP-1 and VCAM-1 in mucosal epithelia and improve inflammatory reaction of AR. CIC-3 plays an important role in the secretion of cytokines and mucosal inflammatory response in AR. RhlL-4 can enhance the secretion of CIC-3, MCP-1 and VCAM-1 in mucosal epithelial cells, especially during the AR process. These enhanced effects of rhlL-4 were significantly suppressed by AHA. The secretions of CIC-3, MCP-1 and VCAM-1 can not be induced obviously by rhlL-2 in mucosal epithelial cells in AR.

CDK4/6 inhibition blocks cancer metastasis through a USP51-ZEB1-dependent deubiquitination mechanism

Tumor metastasis is the most common cause of cancer-related deaths,yet it remains poorly understood.The transcription factor zinc-finger E-box binding homeobox 1(ZEB1)is involved in the epithelial-to-mesenchymal transition(EMT)and plays a pivotal role in tumor metastasis.However,the underlying mechanisms of the posttranslational modification of ZEB1 remain largely unknown.Herein,we demonstrated that specific inhibition of CDK4/6 was able to block tumor metastasis of breast cancer by destabilizing the ZEB1 protein in vitro and in vivo.Mechanistically,we determined that the deubiquitinase USP51 is a bona fide target of CDK4/6.The phosphorylation and activation of USP51 by CDK4/6 is necessary to deubiquitinate and stabilize ZEB1.Moreover,we found a strong positive correlation between the expression of p-RB(an indicator of CDK4/6 activity),p-USP51 and ZEB1 in metastatic human breast cancer samples.Notably,the high expression of p-RB,p-USP51,and ZEB1 was significantly correlated with a poor clinical outcome.Taken together,our results provide evidence that the CDK4/6-USP51-ZEB1 axis plays a key role in breast cancer metastasis and could be a viable therapeutic target for the treatment of advanced human cancers.

Moxifloxacin-induced multiple organ dysfunction possibly related to mutations in several genes involved in drug metabolism pathways

Quinolones have been used to treat various infectious diseases. The addition of fluorine atoms has allowedfor extensive modifications of structures, which has enhanced the spectrum of activities and oral bioavailability of quinolones and resulted in the creation of fluoroquinolones） Although the adverse effects associated with fluoroquinolone antibiotics have decreased considerably and these drugs are generally well tolerated, adverse effects do sometimes occur.