The positive feedback loop of furin and TGFß1 enhances the immune responses of Tregs to hepatocellular carcinoma cells and hepatitis B virus in vitro.

Regulatory T cells (Tregs) can exert immunosuppressive activity. Furin can regulate Treg functions, hepatitis B virus (HBV) persistent infection, and hepatocellular carcinoma (HCC) development. However, it remains unknown whether furin can regulate the immune responses of Tregs to HBV and HCC cells. Here, coculture systems of HBV1.3P-HepG2.3P-HepG2 cells and Tregs transduced with or without lentiviral particles that could overexpress furin or knockdown furin/transforming growth factor ß1 (TGFß1) were established to investigate the regulatory relationship between furin and TGFß1 and the effect of furin/TGFß1 on Treg activity. Also, the effects of furin overexpression or furin/TGFß1 knockdown in Tregs on the immunological activity of effector T cells (Teffs)/cytotoxic T lymphocytes (CTLs) and HBV replication/expression were explored in the coculture system of Teff/CTL, Treg, and HBV1.3P-HepG2 cells. Our results showed that furin expression and TGFß1 secretion were notably increased in Tregs, and Furin and TGFß1 formed a positive feedback loop to activate Tregs in the coculture system of Tregs and HBV1.3P-HepG2 cells. Furin or TGFß1 knockdown in Tregs promoted Teff cell proliferation, stimulated interleukin-2 and interferon-γ secretion, and inhibited HBV replication/gene expression in the coculture system of Teff, Treg, and HBV1.3P-HepG2 cells. Moreover, furin or TGFß1 depletion in Tregs enhanced the killing activity of CTLs against HBV1.3P-HepG2 cells and curbed HBV replication/gene expression in the coculture system of Tregs, CTLs, and HBV1.3P-HepG2 cells. In conclusion, the positive feedback loop of furin and TGFß1 enhanced the immune responses of Tregs to HCC cells and HBV in vitro.

Minimally Invasive Percutaneous Vertebroplasty for Thoracolumbar Instrumented Vertebral Fracture in Patients With Posterior Instrumentation.

BACKGROUND: The traditional treatment for an instrumented vertebral fracture involves removing the loosened pedicle screws and extending the posterior instrumentation cephaladly or caudally. There has been a recent trend of performing minimally invasive fluoroscopy-guided percutaneous vertebroplasty as a salvage procedure. OBJECTIVE: The aim of this study was to compare the outcomes of surgical interventions for instrumented vertebral fracture. STUDY DESIGN: Retrospective assessment. SETTING: All data came from Chang Gung Memorial Hospital, Taiwan. METHODS: We retrospectively reviewed 35 patients with an instrumented vertebral fracture who underwent fluoroscopy-guided percutaneous vertebroplasty (Group I, n = 16) or extension of the posterior instrumentation (Group II, n = 19). Demographic data were recorded. The operating time, amount of intraoperative blood loss, time to postoperative ambulation, and duration of hospital stay were also evaluated. The visual analog scale (VAS) score, kyphotic angle on radiological images, Kirkaldy-Willis functional score, complications, and revision surgery were evaluated at one week and one, 3, 6, and 12 months postoperatively. RESULTS: Group I had a shorter operating time (P < 0.001), less intraoperative blood loss (P < 0.001), earlier postoperative ambulation (P < 0.001), and a shorter hospital stay (P < 0.001). The mean VAS score improved significantly after surgery in both groups (P = 0.001). The postoperative kyphotic angle was better in Group II (P < 0.05). There was no significant between-group difference in the Kirkaldy-Willis functional score at the last follow-up (P = 0.91). There was no significant between-group difference in the need for revision surgery (Group I, n = 4; Group II, n = 5; P = 0.93). LIMITATION: This study is a retrospective cohort. CONCLUSIONS: Minimally invasive fluoroscopy-guided percutaneous vertebroplasty can be used as an alternative to extension of posterior instrumentation for instrumented vertebral fracture. It has several advantages, including a shorter operating time, earlier postoperative ambulation, less blood loss, and a shorter hospital stay. The clinical outcomes of these 2 treatment approaches were similar.

Light control of catechin accumulation is mediated by photosynthetic capacity in tea plant (Camellia sinensis).

BACKGROUND: Catechins are crucial in determining the flavour and health benefits of tea, but it remains unclear that how the light intensity regulates catechins biosynthesis. Therefore, we cultivated tea plants in a phytotron to elucidate the response mechanism of catechins biosynthesis to light intensity changes. RESULTS: In the 250 µmol·m- 2·s- 1 treatment, the contents of epigallocatechin, epigallocatechin gallate and total catechins were increased by 98.94, 14.5 and 13.0% respectively, compared with those in the 550 µmol·m- 2·s- 1 treatment. Meanwhile, the photosynthetic capacity was enhanced in the 250 µmol·m- 2·s- 1 treatment, including the electron transport rate, net photosynthetic rate, transpiration rate and expression of related genes (such as CspsbA, CspsbB, CspsbC, CspsbD, CsPsbR and CsGLK1). In contrast, the extremely low or high light intensity decreased the catechins accumulation and photosynthetic capacity of the tea plants. The comprehensive analysis revealed that the response of catechins biosynthesis to the light intensity was mediated by the photosynthetic capacity of the tea plants. Appropriately high light upregulated the expression of genes related to photosynthetic capacity to improve the net photosynthetic rate (Pn), transpiration rate (Tr), and electron transfer rate (ETR), which enhanced the contents of substrates for non-esterified catechins biosynthesis (such as EGC). Meanwhile, these photosynthetic capacity-related genes and gallic acid (GA) biosynthesis-related genes (CsaroB, CsaroDE1, CsaroDE2 and CsaroDE3) co-regulated the response of GA accumulation to light intensity. Eventually, the epigallocatechin gallate content was enhanced by the increased contents of its precursors (EGC and GA) and the upregulation of the CsSCPL gene. CONCLUSIONS: In this study, the catechin content and photosynthetic capacity of tea plants increased under appropriately high light intensities (250 µmol·m- 2·s- 1 and 350 µmol·m- 2·s- 1) but decreased under extremely low or high light intensities (150 µmol·m- 2·s- 1 or 550 µmol·m- 2·s- 1). We found that the control of catechin accumulation by light intensity in tea plants is mediated by the plant photosynthetic capacity. The research provided useful information for improving catechins content and its light-intensity regulation mechanism in tea plant.

Prognostic Nomogram for Patients with Radical Surgery for Non-Metastatic Colorectal Cancer Incorporating Hematological Biomarkers and Clinical Characteristics.

BACKGROUND: There is a large difference in postoperative survival in patients with non-metastatic colorectal cancer. We aimed to develop nomograms incorporating both hematological biomarkers and clinical characteristics to predict overall survival (OS) in patients with radical surgery for non-metastatic colorectal cancer. METHODS: A retrospective analysis was performed on date from 508 patients who underwent radical resection of colorectal cancer at the Affiliated Tumor Hospital of Guangxi Medical University from December 2011 to December 2015. Simple random sampling was performed by dividing these patients into a training set (n=355) and validation set(n=153), which yielded a 7:3 ratio in the sample sizes between these groups. Based on COX regression analysis of the results from the training cohort, a nomogram was developed to predict the three-year and five-year overall survival rate, and internal verification was also performed. The nomogram prediction accuracy and discriminating ability were evaluated by Harrell's C-index (C-index), calibration curves and were compared with the colorectal cancer TNM staging system. RESULTS: We found that age, degree of differentiation, T stage, N stage, neurological invasion, neutrophils, monocytes, HGB, and LDH were independent risk factors for predicting OS in patients with colorectal cancer. In the training cohort, the C index was 0.796 (95% CI: 0.761-0.831). In the validation cohort, the C index was 0.671 (95% CI: 0.656-0.686).The nomogram showed a stronger predictive ability than did TNM staging. Decision curve analysis showed that the nomogram had value in terms of clinical application. CONCLUSION: Our nomogram combined hematological biomarkers and clinical characteristics and was highly effective in predicting OS in patients with non-metastatic colorectal cancer. Hence, our nomogram may provide a reference tool for clinicians to guide individualized treatment and follow-ups for patients with colorectal cancer.

Percutaneous Balloon Kyphoplasty and Short Instrumentation Compared with Traditional Long Instrumentation for Thoracolumbar Metastatic Spinal Cord Compression.

BACKGROUND: A considerable propotion of patients with cancer got thoracolumbar vertebral metastatic epidural spinal cord compression, which affected their quality of life. Traditional surgical management involves early decompression with concomitant spine stabilization with long instrumentation. However, complications are caused by massive blood loss and long operation time. This study aimed to compare the safety and efficacy of short posterior instrumentation with kyphoplasty and the traditional method for thoracolumbar metastatic epidural spinal cord compression. METHODS: Between January 2004 and December 2015, a retrospective study was conducted on 120 patients with metastatic epidural spinal cord compression from T6 to L5 and divided into 2 groups: short posterior instrumentation with the balloon kyphoplasty group (group I, n = 50) and the long posterior instrumentation group (group II, n = 70). The clinical and radiographic parameters of patients in the 2 groups were compared with a nonrandomized cohort method. Patients were followed up from 3 to 40 months after surgery according to survival time. RESULTS: In group I, the surgery had a mean blood loss of 650 mL and a survival time of 19.1 months. In group II, the surgery had a mean blood loss of 2100 mL and a survival time of 14.14 months. A significant difference in blood loss amount (P = 0.002) was observed. Complications, including deep wound infection, durotomy, and uncontrolled bleeding, were observed in both groups. No postoperative cement-induced neurologic deterioration, implant failure occurred. CONCLUSIONS: Kyphoplasty with short posterior instrumentation is a novel technique that can be performed safely and effectively for the treatment of thoracolumbar metastatic epidural spinal cord compression. Rigid stability, reduced blood loss, and short fixation decrease surgical morbidity of spinal metastasis.

A dielectrophoretic chip with a roughened metal surface for on-chip surface-enhanced Raman scattering analysis of bacteria.

We present an analysis of the results of in situ surface-enhanced Raman scattering (SERS) of bacteria using a microfluidic chip capable of continuously sorting and concentrating bacteria via three-dimensional dielectrophoresis (DEP). Microchannels were made by sandwiching DEP microelectrodes between two glass slides. Avoiding the use of a metal nanoparticle suspension, a roughened metal surface is integrated into the DEP-based microfluidic chip for on-chip SERS detection of bacteria. On the upper surface of the slide, a roughened metal shelter was settled in front of the DEP concentrator to enhance Raman scattering. Similarly, an electrode-patterned bottom layer fabricated on a thin cover-slip was used to reduce fluorescence noise from the glass substrate. Gram positive (Staphylococcus aureus) and Gram negative (Pseudomonas aeruginosa) bacteria were effectively distinguished in the SERS spectral data. Staphylococcus aureus (concentration of 10(6) CFUml) was continuously separated and concentrated via DEP out of a sample of blood cells. At a flow rate of 1 mulmin, the bacteria were highly concentrated at the roughened surface and ready for on-chip SERS analysis within 3 min. The SERS data were successfully amplified by one order of magnitude and analyzed within a few minutes, resulting in the detection of signature peaks of the respective bacteria.