Washed microbiota transplantation for Clostridioides difficile infection: A national multicenter real-world study.

OBJECTIVES: Fecal microbiota transplantation (FMT) has been recommended for the treatment of recurrent Clostridioides difficile infection (CDI). We aimed to evaluate the therapeutic efficacy and safety of washed microbiota transplantation (WMT), a new method of FMT, for CDI across various medical settings. METHODS: This multicenter real-world cohort study included CDI patients undergoing WMT. The primary outcome was the clinical cure rate of CDI within 8 weeks after WMT. Secondary outcomes included the CDI recurrence rate and reduction in total abdominal symptom score (TASS) during the follow-up period. Adverse events related to WMT were recorded. RESULTS: Altogether 90.7% (49/54) of CDI patients achieved clinical cure after treated with WMT. The cure rate was 83.3% for cases with severe and complicated CDI (ScCDI) (n = 30) and 100% for non-ScCDI cases (n = 24) (P = 0.059). No difference was observed in the clinical cure rate between patients with first and recurrent CDI (91.9% vs 88.2%, P = 0.645). One week post-WMT, TASS showed a remarkable decrease compared to that at baseline (P < 0.001). Totally, 8.2% (4/49) of patients suffered CDI recurrence during the follow-up period. A WHO performance score of 4, age ≥65 years, higher TASS score, and higher Charlson comorbidity index score were potential risk factors for efficacy (P = 0.018, 0.03, 0.01, 0.034, respectively). Four (3.8%) transient adverse events related to WMT were observed. CONCLUSIONS: This study emphasizes the attractive value of WMT for CDI. Early WMT may be recommended for CDI, especially for those in serious condition or with complex comorbidities. TRIAL REGISTRATION: ClinicalTrials.gov, no. NCT03895593 (registered on 27 March 2019).

Research on the inhibition for aseptic loosening of artificial joints by Sr-doped calcium polyphosphate (SCPP)in vivo.

Aseptic loosening of artificial joints is the most common complication after artificial joint replacement. Finding the solution to tackle aseptic loosening of artificial joints is a focus in bone and joint surgery research field.In vitrostudies of Sr-doped calcium polyphosphate (SCPP) have found by our team that it could promote osteoblast proliferation and inhibit osteoclast activity, and it has a potential inhibitory effect on aseptic loosening by suppressing the expression of receptor activator of nuclear factor-κ B ligand and improving the expression of OPG. The present study aims to confirm the conclusionin vitroby the mean of animal experiment. The Ti rod prosthesis coated with SCPP, calcium polyphosphate (CPP), and Ultra-high molecular weight polyethylene (UHMWPE were implanted in the femur (the internal surface of bone tunnel was also coated with SCPP, CPP and UHMWPE respectively). Radiography (x-rays, micro-CT), histochemistry (Hematoxylin-eosin staining (HE), methylene blue-acid fuchsin, Von Kossa histological staining), molecular biology (alkaline phosphatase and TRAP5b factors, Mir21-5p and Mir 26a-5p) were performed to analyzed the effects of SCPP within 20 weeks. The Radiography results showed that osteolysis with various severity occurred in all groups, and SCPP group had the mildest osteolysis. Histochemistry results showed that arthritis was milder in SCPP and CPP groups, while the bone formation in SCPP group was most significant. Its bone reconstruction effect was the best as well. The Molecular biology results showed that the bone reconstruction was out-sync in each group. Compared with other groups, the bone resorption occurred at the latest and the bone resorption time was the shortest in experimental animals of SCPP group. All results indicated that SCPP could promote osteoblast activity and bone reconstruction, improve the integration of bone interface between prosthesis and base bone, reduce osteoclast activity and shorten the osteoclast action time at the implantation sitein vivo. Thus, it could postpone or alleviate the occurrence and development of aseptic looseningin vivo. Therefore, SCPP could be a promising material for the construction of artificial joints with the ability to resist aseptic loosening.

The novel circular RNA circ-PGAP3 retards cervical cancer growth by regulating the miR-769-5p/p53 axis.

Cervical cancer (CC) is still an intractable disease that seriously affects women's health. Elucidating its pathogenesis will bring new targets for clinical treatment. Circular RNA (circRNA) is an endogenous RNA that has recently been reported to be closely related to cancer progression and development. In the current study, by performing in silico analysis and qRT-PCR assay, we found a circRNA derived from PGAP3, referred as circ-PGAP3 (hsa_circ_0106800, chr17:37843549-37844086), which was significantly downregulated in CC tissues. Low circ-PGAP3 was closely linked to poor prognosis. And overexpression of circ-PGAP3 significantly reduced CC cell proliferation in vitro and tumor growth in vivo. In terms of mechanism, circ-PGAP3 was transcriptionally elevated by p53, a well-recognized tumor suppressor, and circ-PGAP3 was located in the cytoplasm where sponged miR-769-5p to increase the levels of p53 and its downstream targets. Importantly, the regulatory feedback loop of circ-PGAP3/p53 was also confirmed in vivo. Overall, our data clearly expounded the tumor-inhibiting role of circ-PGAP3 in CC, circ-PGAP3 repressed CC tumorigenesis via regulating the miR-769-5p/p53 axis. Therefore, restoration of circ-PGAP3 may be a promising therapeutic target for this thorny disease.

Anticancer activity of green synthesised AgNPs from Cymbopogon citratus (LG) against lung carcinoma cell line A549.

The present study is designed to analyse the antibacterial and anticancer effects of silver nanoparticles (AgNPs) synthesised from the Cymbopogon citratus, (lemongrass) (LG-AgNPs), which is widely used in ayurvedic drugs for treating various diseases. The LG-AgNPs were synthesised and characterised using ultraviolet (UV) spectroscopy, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD) and transmission electron microscopy (TEM) analysis. The characterised LG-AgNPs was subjected to antimicrobial analysis by disc diffusion method against pathogenic bacteria and fungi. Furthermore, the cytotoxicity and anticancer activity of the LG-AgNPs were assessed with lung alveolar carcinoma cell line A549. Results depict that UV-visible spectra of LG-AgNPs showed strong absorption peak at 435 nm. The XRD study exposed LG-AgNPs crystals, which confirmed with TEM analysis exhibiting particle size ranging between 17 and 25.8 nm. The FTIR spectra recorded peaks at 3347, 2126, 1639, 659, 598 and 553 cm-1. The zone of inhibition study proves the LG-AgNPs possessed both antibacterial and antifungal activities. 3-(4, 5-dimethyl thiazoyl-2-yl)-(2,5-diphenyltetrazolium bromide) results show the cytotoxicity effect of LG-AgNPs in lung cancer cells. It also inhibited the cell migration and invasion at the dose of 25 µg ml-1 by increasing the apoptotic gene expression. The results reveal LG-AgNPs possess anticancer activities, proposing that it may be an alternative drug for allopathic drugs with lots of side effects used in lung cancer treatment.

ISOLATION AND IDENTIFICATION OF AN ENDOPHYTIC FUNGUS PRODUCING PACLITAXEL FROM TAXUS WALLICHIANA VAR MAIREI.

The objective of this study was to isolate endophytic fungi producing paclitaxel from yew for the purpose of paclitaxel manufacture. Surface sterilized bark of Taxus wallichiana var. mairei was used as source material and potato dextrose agar culture medium was used in isolation of endophytic fungi. Fungal cultures were extracted with a mixture of chloroform / methanol (1:1, v/v) and the paclitaxel in the extracts was determined and authenticated with LC-MS. An endophytic fungus that produced paclitaxel was identified by ITS rDNA and 26S D1/D2 rDNA sequencing. The results showed that a total of 435 endophytic fungal strains were isolated from T. wallichiana var. mairei and purified. Only one of these strains produced paclitaxel and it belongs to Fusarium. The paclitaxel productivity in whole PDB culture and that in spent culture medium from this strain is 0.0153 mg/L and 0.0119 mg/L respectively. The paclitaxel content in dry mycelium is 0.27 mg/kg. This isolated endophytic fungus produced paclitaxel at a considerable level and shows potentiality as a producing strain for paclitaxel manufacture after strain improvement.

El objetivo de este estudio fue aislar hongos endofíticos productores de paclitaxel a partir de tejo con el propósito de fabricar paclitaxel. Se utilizó la superficie de la corteza esterilizada de Taxus wallichiana var. mairei como material de origen y dextrosa de patata en medio de cultivo de agar para el aislamiento de hongos endófitos. Los cultivos de hongos se extrajeron con una mezcla de cloroformo / metanol (1:1, v/v) y el paclitaxel en los extractos se determinó y autentificó con LC-MS. Un hongo endófito que produjo paclitaxel fue identificado por su ADNr 26S y secuenciación D1/D2 ADNr. Los resultados mostraron que un total de 435 cepas de hongos endófitos se aislaron y purificarón a partir de T. wallichiana var. mairei. Solo una de estas cepas produce paclitaxel y pertenece a Fusarium. La productividad del cultivo de paclitaxel procedente de esta cepa es 0,0153 mg/L y 0,0119 mg/L, respectivamente. El contenido de paclitaxel en micelio seco es 0,27 mg/kg. Este aislado de hongos endófitos produjo paclitaxel a un nivel considerable y muestra potencial como cepa para la fabricación de paclitaxel después de llevar a cabo una mejora de las cepas.

Downregulation of phosphoglycerate dehydrogenase inhibits proliferation and enhances cisplatin sensitivity in cervical adenocarcinoma cells by regulating Bcl-2 and caspase-3.

Phosphoglycerate dehydrogenase (PHGDH) is the key enzyme of de novo serine biosynthesis. Previous reports have demonstrated that PHGDH plays an important role in some malignancies. However, the biological role of PHGDH in human cervical adenocarcinoma has not been explored. We examined the expression of PHGDH in 54 cervical adenocarcinoma samples by immunohistochemistry and evaluated the association with clinicopathological parameters and prognosis. We performed shRNA transfection to knock down PHGDH gene expression in HeLa cells. A cell proliferation test, cisplatin cytotoxicity test and apoptosis test examined the HeLa cell line after PHGDH knockdown in vitro. In vivo tumorigenesis was assessed using a mouse xenograft model. Moreover, we examined the effects on Bcl-2 and cleaved caspase-3 expression after knockdown of PHGDH and treatment of cisplatin for 48h by Western blot. In this study, we demonstrated that elevated PHGDH expression was found in cervical adenocarcinoma and was associated with tumor size and prognosis. Knocking down PHGDH in HeLa cells significantly inhibited cell proliferation and increased cisplatin chemotherapy sensitivity. Silencing PHGDH resulted in inhibition of tumorigenesis in vivo. Furthermore, PHGDH knockdown reduced Bcl-2 and increased cleaved caspase-3 expression. Collectively, our study indicates the novel roles of PHGDH in cervical adenocarcinoma and identifies PHGDH as a new anticancer target.

[The construction of urine-derived cell lines from patients with spinal muscular atrophy].

Spinal muscular atrophy (SMA) is a common neurodegenerative disease in childhood and infancy, clinically characterized by progressive and symmetric muscular weakness and atrophy. Few effective therapies are available now, and SMA is one of the most common genetic causes of infantile mortality. SMA patient-derived cells are beneficial in basic research on this disease, but the most common model cell, fibroblasts can only be obtained through invasive procedures such as muscle or skin biopsy, which are unwelcome to patients and their families. In this study, fresh urine from SMA patients and healthy controls was collected and centrifuged, and the urine sediment was cultured in vitro. The growth characteristics of urine-derived cells were observed, and the survival of motor neuron (SMN) gene, and the amount and localization of SMN protein in different urine cell lines were investigated. In total, 25 urine cell lines from 11 SMA patients and 14 healthy controls were established. These urine-derived cells expand robustly in vitro with stable cell morphological characteristics. The urine cell lines derived from patients carry the SMN1 gene defect and express a low level of SMN protein, while the intracellular localization of SMN protein is normal. Urine-derived cell culture technology is simple, non-invasive and highly reproducible, a way of obtaining and storing rare cell samples from SMA patients with which to study the pathogenesis of SMA.

Using rhodamine 123 accumulation in CD8 cells as a surrogate indicator to study the P-glycoprotein modulating effect of cepharanthine hydrochloride in vivo.

The purpose of this study was the use of rhodamine 123 (Rho123) accumulation in peripheral blood CD8(+)cells as a surrogate indicator to evaluate the modulating effect of P-glycoprotein (P-gp) inhibitors in the multidrug resistance (MDR) tumor-bearing mouse model. Rho123 was administered to mice, and the fluorescence level in CD8(+) cells was measured. Cepharanthine hydrochloride (CH) and verapamil (VER), two P-gp inhibitors, were administered to mice 1 hour prior to Rho123 administration in vivo or added to peripheral blood 1 hour prior to Rho123 addition ex vivo. The tumor inhibition effect of 5-fluorouracil/adriamycin/cisplatin (FAP) protocol plus CH was also investigated. A concentration- or dose-response relationship was shown between the concentration and dose of CH and Rho123 accumulation or the antitumor activity. In conclusion, the measurement of Rho123 accumulation in CD8(+) cells provides a surrogate assay for the screening of candidate P-gp inhibitors in preclinical trials, and CH is effective in modulating P-gp-mediated MDR in vivo.

Fluorofenidone inhibits Ang II-induced apoptosis of renal tubular cells through blockage of the Fas/FasL pathway.

OBJECTIVES: The present study was designed to investigate the inhibitory effects of fluorofenidone on Ang II-induced apoptosis in renal tubular cells and the related signaling pathway. METHODS: Rat proximal tubular epithelial cells (NRK-52E) were used to examine the anti-apoptosis effects of fluorofenidone. Cell proliferation was assessed by methyl thiazolyl tetrazolium assay. Apoptosis was examined by AO/EB staining and TUNEL assay. The expression of Fas/FasL pathway members, including Fas, FasL, Bax, Bcl-2, Caspase-8, and Caspase-3 was detected by real-time RT-PCR and/or Western blot, respectively. The activity of Caspase-8 and Caspase-3 was detected by spectrophotometry. RESULTS: Fluorofenidone didn't affect the proliferation of NRK-52E cells, but significantly inhibited the apoptosis of NRK-52E cells induced by Ang II. Fluorofenidone significantly reduced Ang II-induced increases in Fas, FasL, Bax, Caspase-8 and Caspase-3 at the mRNA level. Consistent with these observations, fluorofenidone also prevented Ang II-mediated up-regulation of FasL and Bax at the protein level. Additionally, Ang II-induced activation of Caspase-8 and Caspase-3 as well as Ang II-initiated downregulation of Bcl-2 at both mRNA and protein levels was all prevented by fluorofenidone. CONCLUSIONS: Fluorofenidone can inhibit Ang II-induced apoptosis of renal tubular cells through blockage of the Fas/FasL pathway.

Developmental characteristics and aluminum resistance of root border cells in rice seedlings.

The developmental characteristics of root border cells (RBCs) and their role in protection of root apices of rice seedling from Al toxicity were evaluated in two rice (Oryza sativa L.) cultivars differing in Al tolerance. Root elongation and RBCs viability were used as indicators for Al effects. The formation of RBCs and the emergence of the root tip occurred almost simultaneously. Treatment of the root with Al inhibited root elongation and increased Al accumulation in the root tips. Physical removal of RBCs from root tips resulted in a more severe inhibition of root elongation and a higher Al accumulation in the root tips. These effects were more pronounced in the Al-sensitive rice cultivar (II You 6216) than that in the Al-tolerant rice cultivar (II You 838). The relative viability of attached and detached RBCs decreased with increasing Al concentrations. Al also induced a thicker mucilage layer surrounding attached RBCs of both cultivars, and detached RBCs did not. Maintaining the abundant live RBCs encapsulated root tip and enhancing their mucilage secretion, appear to be important in alleviating Al toxicity and in allowing exclusion of Al from the rice root apex.

Silencing of tumor necrosis factor receptor 1 by siRNA in EC109 cells affects cell proliferation and apoptosis.

Tumor necrosis factor receptor 1 (TNFR1) is a membrane receptor able to bind TNF-alpha or TNF-beta. TNFR1 can suppress apoptosis by activating the NF-kappaB or JNK/SAPK signal transduction pathway, or it can induce apoptosis through a series of caspase cascade reactions; the particular effect may depend on the cell line. In the present study, we first showed that TNFR1 is expressed at both the gene and protein levels in the esophageal carcinoma cell line EC109. Then, by applying a specific siRNA, we silenced the expression of TNFR1; this resulted in a significant time-dependent promotion of cell proliferation and downregulation of the apoptotic rate. These results suggest that TNFR1 is strongly expressed in the EC109 cell line and that it may play an apoptosis-mediating role, which may be suppressed by highly activated NF-kappaB.

Bone marrow mesenchymal stem cells upregulate transient outward potassium currents in postnatal rat ventricular myocytes.

Bone marrow mesenchymal stem cell (BMSC) transplantation has been shown to effectively improve cardiac function in experimental animals and patients with myocardial infarction and heart hypertrophy. BMSCs exert potent effects on cardiomyocytes through the inhibition of cardiac apoptosis, the attenuation of cardiac inflammation, etc. However, novel biological actions of BMSCs on cardiomyocytes remain to be explored. The present study was designed to investigate whether BMSCs affect electrophysiological features of neonatal rat ventricular myocytes (NRVMs). BMSCs and NRVMs were indirectly co-cultured at a ratio of 1:10 with a semi-permeable membrane. We found that compared with mono-cultured NRVMs, co-cultured NRVMs exhibited an obvious increase of transient outward potassium current (I(to)), accompanied by significant changes in activation, inactivation and recovery of I(to). Meanwhile, K(V)4.2 mRNA which encodes the channel carrying I(to) was more abundant in co-cultured NRVMs than mono-cultured NRVMs. The increases in basic fibroblast growth factor (bFGF) and insulin growth factor-1 (IGF-1) levels were observed in culture medium of BMSCs. bFGF but not IGF-1 upregulated the K(V)4.2 mRNA expression and enhanced I(to) currents. Taken together, we conclude that BMSCs upregulate I(to) of NRVMs, at least partially, by secreting bFGF that in turn upregulates K(V)4.2 expression and alters the kinetics of I(to).

[P21 expression in renal interstitial fibrosis and regulative effect of enalapril].

OBJECTIVE: To investigate the expression of P21 in renal interstitial fibrosis rats and the effect of enalapril on it. METHODS: Sprague Dawley rats were randomly divided into 3 groups: a sham operation group,a unilateral urethral obstruction group, and an enalapril treatment group. The expression of P21 in renal tubular epithelial cells on the process was detected by immunohistochemistry at different time spots (7, 14, 21 d after UUO, sham-surgery or enalapril treatment). The expression of p21 mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: Seven days after the surgery, significant differences were found in P21 expression between UUO and SOR renal tubular cells. With degree of interstitial fibrosis aggravating, P21 expression increased. Enalapril can inhibit its expression. CONCLUSION: In the kidney of UUO rats, P21 expression increased and enalapril possessed significant inhibitory effects on the procedure. P21 may participate in the pathogenesis of renal tubule-interstitial fibrosis.