Effect of maifan stone on the growth of probiotics and regulation of gut microbiota.

Maifan stone is a kind of mineral medicine in Chinese medicine, which has good adsorption, dissolution, mineralization and biological activity. It has an excellent therapeutic effect on livestock, poultry and aquatic animals suffering from intestinal diseases. This study explored the effect of Maifan stone on the growth ability of Lacticaseibacillus rhamnosus GG (L. rhamnosus GG) and the effect of Maifan stone-L. rhamnosus GG-fermented product on the intestinal inflammation and gut microbiota. We find that Maifan stone can adsorb L. rhamnosus GG to form a carrier bacteria. Maifan stone has the characteristics of acid tolerance and bile salt tolerance and can also improve the activity of L. rhamnosus GG in artificial gastrointestinal juice. The fermented product can reduce the degree of diarrhoea and colon pathology in rats to a certain extent and significantly improve intestinal inflammatory factors and gut microbiota. This study improves the application effect of L. rhamnosus GG in the prevention and treatment of diarrhoea animals and provides a scientific basis for the rational development of Maifan stone resources.

Preferential expansion of CD8+ CD19-CAR T cells postinfusion and the role of disease burden on outcome in pediatric B-ALL.

T cells expressing CD19-specific chimeric antigen receptors (CD19-CARs) have potent antileukemia activity in pediatric and adult patients with relapsed and/or refractory B-cell acute lymphoblastic leukemia (B-ALL). However, not all patients achieve a complete response (CR), and a significant percentage relapse after CD19-CAR T-cell therapy due to T-cell intrinsic and/or extrinsic mechanisms. Thus, there is a need to evaluate new CD19-CAR T-cell products in patients to improve efficacy. We developed a phase 1/2 clinical study to evaluate an institutional autologous CD19-CAR T-cell product in pediatric patients with relapsed/refractory B-ALL. Here we report the outcome of the phase 1 study participants (n = 12). Treatment was well tolerated, with a low incidence of both cytokine release syndrome (any grade, n = 6) and neurotoxicity (any grade, n = 3). Nine out of 12 patients (75%) achieved a minimal residual disease-negative CR in the bone marrow (BM). High disease burden (≥40% morphologic blasts) before CAR T-cell infusion correlated with increased side effects and lower response rate, but not with CD19-CAR T-cell expansion. After infusion, CD8+ CAR T cells had a proliferative advantage over CD4+ CAR T cells and at peak expansion, had an effector memory phenotype with evidence of antigen-driven differentiation. Patients that proceeded to allogeneic hematopoietic cell transplantation (AlloHCT) had sustained, durable responses. In summary, the initial evaluation of our institutional CD19-CAR T-cell product demonstrates safety and efficacy while highlighting the impact of pre-infusion disease burden on outcomes. This trial was registered at www.clinicaltrials.gov as #NCT03573700.

[Berberine alleviates programmed necrosis of metabolic-associated fatty liver disease via activating Nrf2 pathway in mice].

Objective: To investigate the effect of berberine on programmed necrosis of hepatocytes induced by metabolic-associated fatty liver disease (MAFLD) in mice and its related molecular mechanism. Methods: Twenty male C57BL/6N mice were randomly divided into four groups (n=5 in each group): control group (S), fatty liver group (H), berberine group(B), nuclear factor erythroid 2-related factor 2 inhibitor group (Nrf2), and all-trans-retinoic acid (ATRA) group (A). Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), triglycerides (TG), total cholesterol (TC), tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß) concentrations were detected at the end of week 12 to calculate fatty liver index (liver mass/body mass ratio). Liver tissue was stained with HE, Masson and Oil Red O, and SAF score was used to evaluate the degree of liver injury. The expression levels of hepatic programmed necrosis-related proteins, namely receptor-interacting protein kinase 3 (RIPK3), phosphorylated mixed series protease-like domain (p-MLKL) and Nrf2 were detected by Western blot method. One-way ANOVA was used for intragroup comparisons and LSD-t tests were used for intergroup comparisons. Results: Compared with S group, H group serum ALT, AST, LDH, TG, TC, TNF-α, IL-1ß levels and fatty liver index were significantly increased. The liver tissue was filled with vacuolar-like changes and inflammatory cell infiltration. Numerous red lipid droplets were observed with oil red O staining. Collagen fiber hyperplasia was evident with Masson staining. SAF scores (6.60 ± 0.55 and 0.80 ± 0.45) were significantly increased. The expressions of RIPK3 and p-MLKL were up-regulated. Nrf2 level was relatively increased, and the differences were statistically significant (P < 0.05). Compared with H group, berberine intervention group liver biochemical indexes, lipid levels, pro-inflammatory mediator expression, fatty liver index, and SAF score were significantly reduced, and the expression of RIPK3 and p-MLKL were down-regulated, while Nrf2 levels were further increased, and the differences were statistically significant (P<0.05). Compared with B group, treatment with Nrf2 inhibitor had antagonized the protective effect of berberine on fatty liver. Serum ALT, AST, LDH, TG, TC and TNF-α, IL-1ß levels, fatty liver index, and SAF scores were significantly increased and the expressions of RIPK3 and p-MLKL were relatively increased, and the differences were statistically significant (P < 0.05). Conclusion: Berberine can significantly improve the metabolic-associated fatty liver disease injury in mice, and its mechanism is related to activation of Nrf2 and inhibition of programmed necrosis of hepatocytes.

[Effects of protein disulfide isomerase on hyperglycemia and hypoxia/reoxygenation injury in H9c2 cardiomyocytes].

Objective: To explore the effect of protein disulfide isomerase (PDI) in diabetic ischemic heart disease. Methods: We established an in vitro model of high glucose and hypoxia/reoxygenation in H9c2 rat myocardial cells. Cultured cells were divided into four groups: Control, high glucose (HG), hypoxia/reoxygenation (H/R) and HG+H/R. Changes in PDI expression mediated by PDI adenovirus(Ad-PDI) infection and siRNA(PDI-siRNA) transfection in myocardial cells were observed by inverted fluorescence microscopy. We also measured lactate dehydrogenase(LDH) activity and malondialdehyde(MDA) and high molecular weight(HMW)-APN concentrations. PDI, APN, cleaved caspase-3, and glucose regulated protein 78 (Grp78) protein expression were detected. Results: PDI expression was significantly decreased in the HG, H/R and HG+H/R groups compared to the Control group; however, LDH activity[(179.7±10.4) U/L、(218.4±18.4) U/L、(328.2±5.3) U/L vs (91.0±11.0) U/L], MDA concentration[(7.0±0.4) µmol/L、(10.0±1.0) µmol/L、(11.7±1.0) µmol/L vs (4.2±1.8) µmol/L], cleaved caspase-3, and Grp78 expression were increased. Interestingly, APN and HMW-APN expression were decreased [(2.01±0.21) µg/L、(1.64±0.27) µg/L、(1.20±0.14) µg/L vs (2.62±0.12) µg/L, all P<0.05]. Over expression of PDI attenuated high glucose and hypoxia/reoxygenation induced apoptosis and oxidative stress in H9c2 cardiomyocytes(all P<0.05), and simultaneously increased APN and HMW-APN expression [(2.86±0.03) µg/L vs (3.03±0.10) µg/L、(2.06±0.05) µg/L vs (2.31±0.06) µg/L、(1.83±0.07) µg/L vs (1.96±0.11) µg/L、(1.20±0.06) µg/L vs (1.39±0.09) µg/L]. PDI-siRNA transfection increased LDH activity, MDA concentration, and cleaved caspase-3 and Grp78 expression, and decreased APN and HMW-APN expression [(0.75±0.09) µg/L vs (0.59±0.09) µg/L、(0.62±0.04) µg/L vs (0.53±0.05) µg/L、(0.55±0.14) µg/L vs (0.51±0.12) µg/L、(0.48±0.12) µg/L vs (0.35±0.08) µg/L] in response to different treatments in cultured H9c2 cardiomyocytes (all P<0.05). Conclusion: PDI may regulate the expression of APN and HMW-APN, and play an important role in the function of diabetic ischemia-reperfusion cardiomyocytes.

Radiomics of Pediatric Low-Grade Gliomas: Toward a Pretherapeutic Differentiation of BRAF-Mutated and BRAF-Fused Tumors.

BACKGROUND AND PURPOSE: B-Raf proto-oncogene, serine/threonine kinase (BRAF) status has important implications for prognosis and therapy of pediatric low-grade gliomas. Currently, BRAF status classification relies on biopsy. Our aim was to train and validate a radiomics approach to predict BRAF fusion and BRAF V600E mutation. MATERIALS AND METHODS: In this bi-institutional retrospective study, FLAIR MR imaging datasets of 115 pediatric patients with low-grade gliomas from 2 children's hospitals acquired between January 2009 and January 2016 were included and analyzed. Radiomics features were extracted from tumor segmentations, and the predictive model was tested using independent training and testing datasets, with all available tumor types. The model was selected on the basis of a grid search on the number of trees, opting for the best split for a random forest. We used the area under the receiver operating characteristic curve to evaluate model performance. RESULTS: The training cohort consisted of 94 pediatric patients with low-grade gliomas (mean age, 9.4 years; 45 boys), and the external validation cohort comprised 21 pediatric patients with low-grade gliomas (mean age, 8.37 years; 12 boys). A 4-fold cross-validation scheme predicted BRAF status with an area under the curve of 0.75 (SD, 0.12) (95% confidence interval, 0.62-0.89) on the internal validation cohort. By means of the optimal hyperparameters determined by 4-fold cross-validation, the area under the curve for the external validation was 0.85. Age and tumor location were significant predictors of BRAF status (P values = .04 and <.001, respectively). Sex was not a significant predictor (P value = .96). CONCLUSIONS: Radiomics-based prediction of BRAF status in pediatric low-grade gliomas appears feasible in this bi-institutional exploratory study.

CircRNA UBAP2 promotes the progression of ovarian cancer by sponging microRNA-144.

OBJECTIVE: This study aims to elucidate the regulatory effect of circular RNA UBAP2 (circUBAP2) on the progression of ovarian cancer (OC). PATIENTS AND METHODS: Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was used to detect the expressions of circUBAP2, microRNA-144 and CHD2 in OC tissues and adjacent normal tissues. The correlation between the expression levels of circUBAP2 and microRNA-144 with pathological parameters of OC patients was analyzed. Subcellular distribution of circUBAP2 was detected by chromatin fractionation assay. After overexpression of circUBAP2 in OC cells, changes in proliferative and migratory abilities were evaluated by Cell Counting Kit-8 (CCK-8) and transwell assay, respectively. In addition, the Dual-Luciferase reporter gene assay was used to verify the binding of circUBAP2 and microRNA-144, and the binding of CHD2 to microRNA-144. RESULTS: QRT-PCR results showed that circUBAP2 was highly expressed in OC tissues, and its expression was negatively correlated with TMN stage and five-year survival of OC patients. CircUBAP2 was mainly distributed in the cytoplasm. Overexpression of circUBAP2 significantly promoted the proliferative and migratory abilities of OC cells. The Dual-Luciferase reporter gene assay demonstrated that circUBAP2 could bind to microRNA-144. Meanwhile, circUBAP2 negatively regulated microRNA-144 expression in OC cells. Besides, the promotive effects of circUBAP2 on the proliferation and migration of OC cells were reversed by microRNA-144 overexpression. MicroRNA-144 was lowly expressed in OC tissues, which was negatively correlated with TNM stage of OC patients. The Dual-Luciferase reporter gene assay confirmed the binding condition between CHD2 and microRNA-144. CHD2 expression was negatively regulated by microRNA-144 in OC cells. Moreover, CHD2 could bind to microRNA-144 and partially inhibited its activity, thereby promoting the proliferative and migratory abilities of OC cells. CONCLUSIONS: CircUBAP2 promotes the progression of ovarian cancer by adsorbing microRNA-144.

Marsupialisation for the treatment of unicystic ameloblastoma of the mandible: a long-term follow up of 116 cases.

Unicystic ameloblastoma is a unique histopathological type of ameloblastoma, and treatment is controversial. Marsupialisation is effective in reducing the size of cystic lesions and their complications. We have retrospectively analysed the clinical, histopathological, and prognostic data of affected patients who were treated by marsupialisation between 2003 and 2013 in three Chinese hospitals. Our aim was to evaluate the effects and prognosis, and the factors associated with outcome. A total of 116 patients with mandibular unicystic ameloblastomas were included, and 74, 26, and 16 patients were histopathologically classified as being luminal, intraluminal, and mural subtypes, respectively. Most responded well to marsupialisation, with an overall recurrence rate of 12%. Resorption of the root (p<0.001), perforation of the cortical bone (p=0.005), and histopathological subtype (p=0.013) were the main factors that predicted the outcome. Perforation of the cortical bone was the only reliable predictor of recurrence (p<0.001). Disease-free survival function curves indicated that patients with the mural subtype were at a higher risk of recurrence than patients with the other two subtypes (p=0.003). Poor outcomes of marsupialisation were treated surgically and, to date, no subsequent recurrences have been reported. Marsupialisation is effective for these patients, with a recurrence rate similar to that of radical treatment. The outcomes can be predicted using characteristics of the lesion such as resorption of the root, perforation of the cortical bone, and histopathological subtypes. However, additional studies are required to corroborate these findings.

Comparison of surgical techniques for benign parotid tumours: a multicentre retrospective study.

This study aimed to compare the outcomes of three surgical techniques for the treatment of patients with benign parotid tumours: superficial parotidectomy (SP; group 1), partial superficial parotidectomy (PSP; group 2), and ultrasonic scalpel-assisted minimal extracapsular dissection (US-MECD; group 3). Groups 1 and 2 received the conventional surgical technique, while group 3 underwent surgery with an ultrasonic scalpel. A total of 281 patients treated during 2012-2016 were included: 98 in group 1, 91 in group 2, and 92 in group 3. The mean surgical time and blood loss during surgery, as well as drainage time and amount, were significantly lower for US-MECD (P<0.01). The great auricular nerve and parotid fascia were both preserved with US-MECD (P<0.01), while the rate of capsule rupture with US-MECD was slightly higher than in the other groups (P>0.05). There was less transient facial nerve paralysis and Frey syndrome with US-MECD (P<0.01). No significant difference in wound infection, sialocele, or permanent facial nerve paralysis was observed among the three groups. Patients enrolled during 2012-2013 were selected to evaluate the recurrence rates, and no statistically significant differences were found among the groups. In conclusion, US-MECD showed similar effectiveness and fewer side effects than SP and PSP. The long-term effects of the new technique require further study.

CXCL13 inhibition induce the apoptosis of MDA-MB-231 breast cancer cells through blocking CXCR5/ERK signaling pathway.

OBJECTIVE: Treatment of the high-risk triple negative breast cancer (TNBC) is a critical clinical challenge. Here we aimed to explore a novel strategy for TNBC treatment by blocking the tumor-associated chemokine CXCL13 in the MDA-MB-231 TNBC cells. MATERIALS AND METHODS: MDA-MB-231 cells were treated with anti-CXCL13 antibodies (inhibition group), or phosphate-buffered saline (PBS) (control group), followed by determining the levels of interleukin-1 (IL-1), tumor necrosis factor-alpha (TNF-α) and transforming growth factor beta-1 (TGF-ß1) with enzyme-linked immunosorbent assay (ELISA). The effects of CXCL13 inhibition on cell proliferation and apoptosis were assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and flow cytometry, respectively. Quantitative Real Time-PCR (qRT-PCR) and Western blot were used to compare the levels of CXCL13, CXCR5, extracellular signal-regulated kinase (ERK). The levels of cyclin D1 and cleaved caspase-9 were detected by Western blot. RESULTS: The levels of IL-1, TNF-α and TGF-ß1 in MDA-MB-231 cells treated with anti-CXCL13 antibodies were significantly downregulated (p<0.05). Meanwhile, CXCL13 blockade decreased the cell proliferation and increased the apoptosis rate of MDA-MB-231 cells. The inhibition of CXCL13 led to marked reduction in CXCL13 and CXCR5 mRNA and an increase in ERK mRNA. The inhibition of CXCL13 resulted in the downregulation of CXCL13, CXCR5, p-ERK/ERK, cyclin D1 and upregulation of cleaved caspase-9 proteins. CONCLUSIONS: CXCL13 blockade effectively suppressed the proliferation of MDA-MB-231 cells by promoting cell apoptosis. This effect is presumably associated with the downregulation of CXCL13 and suppression of the CXCR5/ERK signaling pathway.

Metformin enhances inhibitive effects of carboplatin on HeLa cell proliferation and increases sensitivity to carboplatin by activating mitochondrial associated apoptosis signaling pathway.

OBJECTIVE: Cervical cancer has become the fourth most common cancer in developing countries. This study aimed to investigate anti-tumor effects of Metformin combining with carboplatin in cervical cell line, HeLa cell. MATERIALS AND METHODS: Human cervical cancer cell line, HeLa cell, was treated with Metformin (5 mmol/l or 10 mmol/l) or/and carboplatin (25 mg/l or 50 mg/l) at different final concentrations, and divided into 8 groups. 3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyltetrazolium bromide (MTT) assay was used to evaluate cell viability. Acridine orange/ethidium bromide (AO/EB) staining was used to examine nuclear fragments and cell apoptosis. Annexin V/propidium iodide (PI) staining was employed to detect apoptosis of HeLa cells. Mitochondrial membrane potential of the HeLa cells was evaluated by staining with 5,5,6,6-tetrachloro-1,1,3,3-tetraethylbenzimidazolylcarbocyanine iodide (JC-1) reagent. RESULTS: MTT results showed that Metformin combining carboplatin significantly reduced HeLa cell viability compared to that of no-drug treatment group (p<0.05). Metformin combining carboplatin significantly increased the amounts of nuclear fragments compared to that of no-drug treatment group (p<0.05). The flow cytometry assay results indicated that Metformin combining carboplatin significantly enhanced the apoptotic rates compared to that of no-drug treatment group (p<0.05). The JC-1 staining findings illustrated that Metformin combining carboplatin significantly decreased the mitochondrial membrane potential compared to that of no-drug treatment group (p<0.05). CONCLUSIONS: Metformin enhanced the inhibitive effects of carboplatin on HeLa cell proliferation. Metformin increased the sensitivity of HeLa cell to the treatment of Carboplatin by activating mitochondrial-associated apoptosis signaling pathway.

Compensatory Strategies That Reduce Knee Extensor Demand During a Bilateral Squat Change From 3 to 5 Months Following Anterior Cruciate Ligament Reconstruction.

Background Decreased extensor moments in the surgical knee during bilateral squats can persist beyond 1 year following anterior cruciate ligament reconstruction (ACLR). This is accomplished using interlimb and intralimb compensations. Objectives This study sought to assess loading during squatting longitudinally, 3 and 5 months post ACLR, and to determine the extent to which interlimb and intralimb compensations contribute to reduced knee extensor moments. Methods In this controlled, longitudinal laboratory study, 11 individuals (4 male) underwent 3-D motion analysis of a squat at 3 and 5 months post ACLR. A repeated-measures multivariate analysis of variance (limb by time) assessed differences in peak knee and hip flexion angles, knee extensor moment, vertical ground reaction force, and hip-to-knee extensor moment ratio. Stepwise linear regression analysis was used to determine the contribution of interlimb (between-limb vertical ground reaction force ratio) and intralimb (within-surgical-limb hip-to-knee moment ratio) compensations to the between-limb knee extensor moment ratio. Results A significant effect of limb was observed for knee flexion angle, knee extensor moment, vertical ground reaction force, and hip-to-knee extensor moment ratio, while a significant effect of time was observed for knee extensor moment and hip-to-knee extensor moment ratio. At 3 months, the vertical ground reaction force ratio and hip-to-knee extensor moment ratio predicted the knee extensor moment ratio (R2 = 0.854, P<.001). At 5 months, the hip-to-knee extensor moment ratio predicted the knee extensor moment ratio (R2 = 0.584, P = .006). Conclusion Individuals used interlimb and intralimb compensations to reduce the knee extensor moment of the surgical limb at 3 months post ACLR. Similar reductions in the knee extensor moment at 5 months were accomplished with only intralimb compensations. J Orthop Sports Phys Ther 2018;48(9):713-718. Epub 12 Jun 2018. https://doi.org/10.2519/jospt.2018.7977.

Characterizing knee loading asymmetry in individuals following anterior cruciate ligament reconstruction using inertial sensors.

Limitations in the ability to identify knee extensor loading deficits during gait in individuals following anterior cruciate ligament reconstruction (ACLr) may underlie their persistence. A recent study suggested that shank angular velocity, directly output from inertial sensors, differed during gait between individuals post-ACLr and controls. However, it is not clear if this kinematic variable relates to knee moments calculated using joint kinematics and ground reaction forces. Heel rocker mechanics during loading response of gait, characterized by rapid shank rotation, require knee extensor control. Measures of shank angular velocity may be reflective of knee moments. This study investigated the relationship between shank angular velocity and knee extensor moment during gait in individuals (n=19) 96.7±16.8days post-ACLr. Gait was assessed concurrently using inertial sensors and a marker-based motion system with force platforms. Peak shank angular velocity and knee extensor moment were strongly correlated (r=0.75, p<0.001) and between limb ratios of angular velocity predicted between limb ratios of extensor moment (r(2)=0.57, p<0.001) in the absence of between limb differences in spatiotemporal gait parameters. The strength of these relationships indicate that shank kinematic data offer meaningful information regarding knee loading and provide a potential alternative to full motion analysis systems for identification of altered knee loading following ACLr.

miR-218 tissue expression level is associated with aggressive progression of gastric cancer.

The aim of the present study was to investigate the clinical significance of microRNA-218 (miR-218) in gastric cancer. We enrolled 112 patients having undergone surgery for gastric cancer between May 2008 and June 2014. Expression of miR-218 was determined by real-time quantitative reverse transcription-polymerase chain reaction. Survival curves were plotted using the Kaplan-Meier method and compared by the log-rank test. We found that miR-218 expression was significantly downregulated in gastric cancer tissues compared to adjacent normal tissues (P < 0.001). Low miR-218 expression was significantly associated with tumor differentiation (P < 0.001), depth of tumor invasion (P = 0.006), and tumor node metastasis stage (P < 0.001). Kaplan-Meier survival analysis revealed that patients with low miR-218 levels showed significantly lower 5-year overall survival than those demonstrating high expression (P = 0.04). Multivariate Cox regression analyses indicated that low miR-218 expression constitutes an independent molecular biomarker for prediction of poor overall survival of gastric cancer patients (hazard ratio = 3.187, 95% confidence interval = 1.551-8.365, P = 0.037). In conclusion, miR-218 was remarkably downregulated in gastric cancer tissues and may serve as a prognostic biomarker for patients suffering from this disease.

Local administration of AAV-DJ pseudoserotype expressing COX2 provided early onset of transgene expression and promoted bone fracture healing in mice.

We have previously obtained compelling proof-of-principle evidence for COX2 gene therapy for fracture repair using integrating retroviral vectors. For this therapy to be suitable for patient uses, a suitable vector with high safety profile must be used. Accordingly, this study sought to evaluate the feasibility of AAV as the vector for this COX2 gene therapy, because AAV raises less safety issues than the retroviral vectors used previously. However, an appropriate AAV serotype is required to provide early increase in and adequate level of COX2 expression that is needed for fracture repair. Herein, we reported that AAV-DJ, an artificial AAV pseudoserotype, is highly effective in delivering COX2 gene to fracture sites in a mouse femoral fracture model. Compared with AAV-2, the use of AAV-DJ led to ~5-fold increase in infectivity in mesenchymal stem cells (MSCs) and provided an earlier and significantly higher level of transgene expression at the fracture site. Injection of this vector at a dose of 7.5 × 10(11) genomic copies led to high COX2 level at the fracture site on day 3 after injections and significantly promoted fracture union at 21 days, as analyzed by radiography and µ-CT. The therapeutic effect appears to involve enhanced osteoblastic differentiation of MSCs and remodeling of callus tissues to laminar bone. This interpretation is supported by the enhanced expression of several key genes participating in the fracture repair process. In conclusion, AAV-DJ is a promising serotype for the AAV-based COX2 gene therapy of fracture repair in humans.

Protective effect of Berberine pretreatment in hepatic ischemia/reperfusion injury of rat.

BACKGROUND: Berberine (BBR) has been demonstrated to protect against hepatic ischemia/reperfusion (I/R) injury. However, the exact mechanism is largely unknown. In the present study, we examined the role of phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) and mammalian target of rapamycin (mTOR) in the protective effect of BBR on hepatic I/R-mediated apoptosis in rats. METHODS: Adult male Sprague-Dawley rats were assigned randomly to groups of sham, ischemia/reperfusion (I/R), I/R+DMSO (vehicle) and I/R+BBR (100 mg/kg/d, 2 weeks). The hepatic cold ischemia model was established by perfusing the liver with heparinized cold saline through the portal vein for 20 minutes. The liver function and oxidative stress level were measured by biochemical and histopathologic examinations. The apoptotic rate was determined by terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL) assay. The expression of Bcl-2, Bax, caspase-3, and the phosphorylation of Akt and mTOR were assayed by Western blot analysis and immunohistochemistry. RESULTS: Compared with the I/R group, BBR dramatically attenuated the histopathologic damage, restored the liver function, and decreased the oxidative stress level. Simultaneously, BBR significantly ameliorated apoptosis by decreasing the apoptotic rate, increasing the Bcl-2/Bax ratio and inhibiting cleaved caspase-3 expression in rats subjected to hepatic I/R. The expression of p-Akt were effectively upregulated with the inhibited expression of p-mTOR. CONCLUSION: Our study reveals that BBR preconditioning protects against hepatic I/R partly by reducing apoptosis, which is possibly involved with the modulation of the PI3K/Akt/mTOR signaling pathway.

First Report of Fusarium solani Causing Root Rot on Coptis chinensis in Southwestern China.

China is the world's largest producer country of coptis (Coptis chinensis), the rhizomes of which are used in traditional Chinese medicine. Since 2008, however, root rot symptoms, including severe necrosis and wilting, have been observed on coptis plants in Chongqing, southwestern China. Of the plants examined from March 2011 to May 2013 in 27 fields, 15 to 30% were covered with black necrotic lesions. The leaves of infected plants showed wilt, necrotic lesions, drying, and death. The fibrous roots, storage roots, and rhizomes exhibited brown discoloration and progressive necrosis that caused mortality of the infected plants. Infected plants were analyzed to identify the causal organism. Discoloration of the internal vascular and cortical tissues of the rhizomes and taproots was also evident. Symptomatic taproots of the diseased coptis were surface sterilized in 1% sodium hypochlorite for 2 min, rinsed in sterile distilled water for 2 min, and then air-dried in sterilized atmosphere/laminar flow. Small pieces of disinfested tissue (0.3 cm in length) were transferred to petri dishes containing potato dextrose agar (PDA) supplemented with 125 µg ml-1 streptomycin sulfate and 100 µg ml-1 ampicillin, and incubated for 5 days at 25°C with a 12-h photoperiod. Four distinct species of fungal isolates (HL1 to 4) derived from single spores were isolated from 30 plants with root rot symptoms collected from the study sites. To verify the pathogenicity of individual isolates, healthy coptis plants were inoculated by dipping roots into a conidial suspension (106 conidia/ml) for 30 min (15 plants per isolate), as described previously (1). Inoculated plants were potted in a mixture of sterilized quartz sand-vermiculite-perlite (4:2:1, v/v) and incubated at 25/18°C and 85 to 90% relative humidity (day/night) in a growth chamber with a daily 16-h photoperiod of fluorescent light. Plants dipped in sterile distilled water were used as controls. After 15 days, symptoms similar to those observed in the field were observed on all plants (n = 15) that were inoculated with HL1, but symptoms were not observed on plants inoculated with HL2, HL3, and HL4, nor on control plants. HL1 was re-isolated from symptomatic plants but not from any other plants. Morphological characterization of HL1 was performed by microscopic examination. The septate hyphae, blunt microconidia (2 to 3 septa) in the foot cell and slightly curved microconidia in the apical cell, and chlamydospores were consistent with descriptions of Fusarium solani (2). The pathogen was confirmed to be F. solani by amplification and sequencing of the ribosomal DNA internal transcribed spacer (rDNA-ITS) using the universal primer pair ITS4 and ITS5. Sequencing of the PCR product revealed a 99 to 100% similarity with the ITS sequences of F. solani in GenBank (JQ724444.1 and EU273504.1). Phylogenetic analysis (MEGA 5.1) using the neighbor-joining algorithm placed the HL1 isolate in a well-supported cluster (97% bootstrap value based on 1,000 replicates) with JQ724444.1 and EU273504.1. The pathogen was thus identified as F. solani based on its morphological and molecular characteristics. To our knowledge, this is the first report of root rot of coptis caused by F. solani in the world. References: (1) K. Dobinson et al. Can. J. Plant Pathol. 18:55, 1996. (2) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Oxford, 2006.

Radiological findings in 210 paediatric patients with viral pneumonia: a retrospective case study.

OBJECTIVE: The objective of this study was to assess the radiological presentations of different types of viral pneumonia in children. METHODS: Nasopharyngeal swab specimens and bronchial aspirate samples from children with acute respiratory infections were obtained and tested for influenza B, adenovirus, respiratory syncytial virus and parainfluenza (Types 1, 2 and 3) by direct immunofluorescence assay, or for influenza A (Subtype H1N1) by quantitative real-time polymerase chain reaction. The chest radiographs of the 210 confirmed cases of viral pneumonia were analysed retrospectively by two independent radiologists for the identification, characterisation and description of the distribution of imaging abnormalities. The cases were divided into six groups on the basis of confirmed causative viral agent, and radiographic findings were compared, analysed and presented. RESULTS: The abnormal chest radiograph findings consisted of bilateral patchy areas of consolidation (n=133), interstitial lung disease (n=33), diffuse areas of air space consolidation (n=29) and lobar consolidation (n=15). The abnormalities were distributed bilaterally in 195 cases and observed more frequently in the lower zones than in other regions. The radiological findings varied significantly among the six groups (p=0.0050). Pairwise comparison showed significant difference between influenza A (H1N1) and adenovirus (p=0.0031) only. CONCLUSION: The predominant radiological finding in paediatric viral pneumonia was bilateral patchy areas of consolidation. The radiological findings differed significantly only between adenovirus and influenza A pneumonia. The diagnosis of the specific causative organism requires laboratory confirmation.

A critical role for NMDA receptors in parvalbumin interneurons for gamma rhythm induction and behavior.

Synchronous recruitment of fast-spiking (FS) parvalbumin (PV) interneurons generates gamma oscillations, rhythms that emerge during performance of cognitive tasks. Administration of N-methyl-D-aspartate (NMDA) receptor antagonists alters gamma rhythms, and can induce cognitive as well as psychosis-like symptoms in humans. The disruption of NMDA receptor (NMDAR) signaling specifically in FS PV interneurons is therefore hypothesized to give rise to neural network dysfunction that could underlie these symptoms. To address the connection between NMDAR activity, FS PV interneurons, gamma oscillations and behavior, we generated mice lacking NMDAR neurotransmission only in PV cells (PV-Cre/NR1f/f mice). Here, we show that mutant mice exhibit enhanced baseline cortical gamma rhythms, impaired gamma rhythm induction after optogenetic drive of PV interneurons and reduced sensitivity to the effects of NMDAR antagonists on gamma oscillations and stereotypies. Mutant mice show largely normal behaviors except for selective cognitive impairments, including deficits in habituation, working memory and associative learning. Our results provide evidence for the critical role of NMDAR in PV interneurons for expression of normal gamma rhythms and specific cognitive behaviors.

Human steroidogenic factor-1 (hSF-1) regulates progesterone biosynthesis and growth of ovarian surface epithelial cancer cells.

The majority of cancers derived from ovarian surface epithelial (OSE) cells are lethal. Estrogens promote proliferation of OSE cells, whereas progesterone inhibits proliferation and promotes apoptosis of OSE cells. Human steroidogenic factor-1 (hSF-1) induction of the steroidogenic acute regulatory protein (StAR) gene, and the steroidogenic enzymes CYP11A1 and HSD3B2 is central to progesterone biosynthesis. Whereas hSF-1 and StAR are expressed in human ovarian surface epithelial (HOSE) cells, hSF-1 and StAR protein were not expressed in a panel of malignant ovarian cancer cell lines (SKOV-3, BG-1, and Caov-3), and in human OSE cells immortalized by SV40 large T antigen (IOSE-121). Transient expression of hSF-1 in SKOV-3 cells activated the expression of StAR, p450scc and 3betaHSD-II mRNAs, and induced progesterone biosynthesis. Additionally, hSF-1 suppressed proliferation and promoted apoptosis of SKOV-3 cells and suppressed SKOV-3 cell growth induced by ERalpha and estradiol. These findings suggest that hSF-1 is central to progesterone biosynthesis in OSE cells. Human SF-1 may decrease OSE cancer cell numbers directly by apoptosis, and indirectly by opposing estradiol-induced proliferation. These findings are consistent with the hypothesis, that down-regulation of hSF-1 contributes to progression of ovarian epithelial cancers.