Accurate Prediction of Metachronous Liver Metastasis in Stage I-III Colorectal Cancer Patients Using Deep Learning With Digital Pathological Images.

Objectives: Metachronous liver metastasis (LM) significantly impacts the prognosis of stage I-III colorectal cancer (CRC) patients. An effective biomarker to predict LM after surgery is urgently needed. We aimed to develop deep learning-based models to assist in predicting LM in stage I-III CRC patients using digital pathological images. Methods: Six-hundred eleven patients were retrospectively included in the study and randomly divided into training (428 patients) and validation (183 patients) cohorts according to the 7:3 ratio. Digital HE images from training cohort patients were used to construct the LM risk score based on a 50-layer residual convolutional neural network (ResNet-50). An LM prediction model was established by multivariable Cox analysis and confirmed in the validation cohort. The performance of the integrated nomogram was assessed with respect to its calibration, discrimination, and clinical application value. Results: Patients were divided into low- and high-LM risk score groups according to the cutoff value and significant differences were observed in the LM of the different risk score groups in the training and validation cohorts (P<0.001). Multivariable analysis revealed that the LM risk score, VELIPI, pT stage and pN stage were independent predictors of LM. Then, the prediction model was developed and presented as a nomogram to predict the 1-, 2-, and 3-year probability of LM. The integrated nomogram achieved satisfactory discrimination, with C-indexes of 0.807 (95% CI: 0.787, 0.827) and 0.812 (95% CI: 0.773, 0.850) and AUCs of 0.840 (95% CI: 0.795, 0.885) and 0.848 (95% CI: 0.766, 0.931) in the training and validation cohorts, respectively. Favorable calibration of the nomogram was confirmed in the training and validation cohorts. Integrated discrimination improvement and net reclassification index indicated that the integrated nomogram was superior to the traditional clinicopathological model. Decision curve analysis confirmed that the nomogram has clinical application value. Conclusions: The LM risk score based on ResNet-50 and digital HE images was significantly associated with LM. The integrated nomogram could identify stage I-III CRC patients at high risk of LM after primary colectomy, so it may serve as a potential tool to choose the appropriate treatment to improve the prognosis of stage I-III CRC patients.

Effect of Jinzhen granule on two coronaviruses: The novel SARS-CoV-2 and the HCoV-229E and the evidences for their mechanisms of action.

BACKGROUND: Although severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and human coronavirus 229E (HCoV-229E) pose a huge threat to human public health, no specific treatment is available. Jinzhen granule (JZ) is a traditional eight ingredients-Chinese medicine with prominent efficacy for treating viral-induced diseases. However, little is known about the antiviral effect and mechanism of JZ against SARS-CoV-2 and HCoV-229E. PURPOSE: This study aimed to reveal the antiviral effects of JZ against SARS-CoV-2 and HCoV-229E, and to further explore the underlying mechanisms regulating the host immune response. METHODS: The chromatographic separation of JZ was performed using a Shimadzu analytical high-performance liquid chromatograph with UV detection and Alltech ELSD 2000ES. We conducted cytopathic effect (CPE) and plaque reduction assays to evaluate the antiviral effect of JZ. A lethal human angiotensin converting enzyme 2 (hACE2) transgenic mouse model of SARS-CoV-2 was established to determine the protective effect of JZ on mortality and lung virus titers. Real-time quantitative PCR assays were used to analyze the expression of proinflammatory cytokines in vitro and in vivo. Western blotting was further performed to determine the activities on regulating the nuclear factor kappa B (NF-κB)/MAPK pathway. Finally, mitochondrial membrane potential assays, flow cytometry analysis and western blotting were used to assess the anti-apoptotic potency toward HCoV-229E infection. RESULTS: The results showed that 13 chemical components were identified and five peaks were determined and quantitated (gallic acid 1.97 mg/g, baicalin 20.69 mg/g, glycyrrhizic acid 4.92 mg/g, hyodeoxycholic acid 4.86 mg/g, cholic acid 4.07 mg/g). We found that JZ exerted inhibitory potency against SARS-CoV-2 and HCoV-229E in vitro by using CPE and plaque reduction assays, and it was further found that JZ protected mice infected by SARS-CoV-2 from death and inhibited lung virus titers. JZ also significantly decreased the induction of inflammatory cytokines (IL-1α, IL-6, CCL-5 and MIP-1ß), similar to the observed in vitro effect. Moreover, JZ suppressed the release of inflammatory cytokines in vitro and it decreased the protein expression of p-p38 MAPK, p-JNK, p-NF-κB p65 and p-IκBα induced by HCoV-229E and increased the expression of IκBα. Notably, JZ significantly protected HCoV-229E-infected Huh-7 cells from mitochondrial damage and decreased apoptotic cells. The activation of the mitochondria-mediated apoptotic pathway was inhibited by JZ, as shown by the reduced expression of cleaved caspase-9, caspase-3 and p-PARP. CONCLUSIONS: In conclusion, JZ (gallic acid 1.97 mg/g, baicalin 20.69 mg/g, glycyrrhizic acid 4.92 mg/g, hyodeoxycholic acid 4.86 mg/g, cholic acid 4.07 mg/g) exhibited antiviral activities against SARS-CoV-2 and HCoV-229E by regulating the NF-κB/MAPK pathway and the mitochondria-mediated apoptotic pathway. These findings demonstrated the efficacy of JZ against CoVs and suggested JZ treatment as a novel clinical therapeutic strategy for COVID-19.

Reducing the effects of control materials based on interchangeability of estimates of day-to-day imprecision between commercial control materials and serum samples.

BACKGROUND: Reduce the effects in the storage-and-thawing process of commercial control materials based on their interchangeability evaluation. METHODS: Seven assays-anti-streptolysin O, complement 3, carcinoembryonic antigen, urea, ferritin, total bilirubin, and glucose-were selected. Commercial control materials and serum samples with similar concentrations were chosen as samples. The experiment was carried out in three stages. In the first stage, the assays with statistical differences in imprecision were screened. In the second stage, two specimens were sealed with parafilm and frozen at -80°C and thawed in the water bath, and the imprecision differences were compared again. Finally, the effective means to reduce the effects were included in the standard operating procedure to repeat confirmation. RESULTS: In the first stage, there was only a statistical difference (p < 0.05) in the imprecision of glucose and total bilirubin between two specimens, and the imprecision of control materials was higher than the serum samples. In the second stage, glucose imprecision was not statistically different (p > 0.05) and lower than in the first stage. In the third stage, the methods from the second stage were confirmed to be effective at reducing control material effects. CONCLUSION: Finding variation factors and confirming and standardizing the measures will help lessen commercial control material effects.

Snare-assisted endoscopic resection of gastric subepithelial tumors originating from the muscularis propria layer: a multicenter study.

BACKGROUND AND AIM: Although successful endoscopic resection of gastric subepithelial tumors (SETs) originating from the muscularis propria (MP) layer has been frequently reported, it requires a relatively complicated technique and has a high perforation rate. In this retrospective study, we evaluated the efficacy and safety of the snare-assisted endoscopic resection (SAER) method which is performed using a snare and insulated-tip (IT) knife via a single-channel endoscope to reduce the perforation rate. METHODS: In this study, fifty-six patients with gastric SETs originating from the MP layer treated by the SAER method at three institutions between July 2017 and December 2017 were reviewed. The procedure involved multiple steps as shown in Fig. 2. Data were obtained on demographics, SET features, histopathological diagnoses, procedure time, en bloc resection rate, R0 resection (negative margins) status, and adverse events. RESULTS: Endoscopic resection was successfully performed in all patients. The median overall procedure time was 43.5 min (range 26-106 min). The mean size of resected specimens was 19.73 mm (range 10-33 mm). The overall rate of en bloc resection was 96.4% (54/56). In addition, the perforation rate was 7.1% (4/56), and defects in the stomach wall were very small and easily closed using metallic clips. No postprocedural bleeding occurred in any case. CONCLUSIONS: The SAER method is an effective, safe, less costly technique for the removal of some gastric SETs originating from the MP layer with an appropriate size.

Novel nomogram with microvascular density in the surgical margins can accurately predict the risk for anastomotic leakage after anterior resection for rectal cancer.

BACKGROUND: Anastomotic leakage (AL) is a serious complication after anterior resection. The purpose of this study was to determine the role of microvascular density (MVD) in AL and to develop a nomogram to accurately predict AL. METHODS: This study retrospectively enrolled 477 consecutive patients who underwent anterior resection for rectal cancer from January 2011 to January 2019. Tissue samples of the resection margins were assessed for MVD. Univariate and multivariate regression analyses were used to identify the risk factors for AL. RESULTS: The incidence of clinical AL was 6.7%. MVD in the distal margin was associated with AL (P < .001). Univariate and multivariate regression analysis identified the following variables as independent risk factors for AL: preoperative albumin ≤35 g/L (odds ratio [OR] = 2.511), neoadjuvant treatment (OR = 3.560), location of tumor ≤7 cm (OR = 3.381), blood loss ≥100 mL (OR = 2.717), and MVD in the distal margin ≤20 (OR = 4.265). Then, a nomogram including these predictors was developed. The nomogram showed good discrimination (AUC = 0.816) and calibration (concordance index = 0.816). The decision curve analysis demonstrated that the nomogram was clinically useful. CONCLUSIONS: MVD in the distal margin is closely associated with AL. The nomogram can be used for individualized prediction of AL after anterior resection for patients with rectal cancer.

Inhibition of AKT enhances the anti-cancer effects of Artemisinin in clear cell renal cell carcinoma.

Given studies have shown that Artemisinin (ART) reduces cancer cell proliferation, migration, invasion, tumorigenesis and metastasis. In this study, we evaluated the roles of ART in clear cell renal cell carcinoma (ccRCC) progression. We measured the eff ;ects of ART on cancer cell proliferation, colony formation, migration, invasion and tumorigenesis. CCK-8 assay demonstrated that ART inhibited cell growth with IC50 values 31.30 ±â€¯0.73 µM in UMRC-2 and 23.97 ±â€¯0.92 µM in CAKI-2, respectively. Colony formation assay shown that ART inhibited cell colony formation. Transwell migration and invasion assay shown that ART inhibited RCC migration and invasion. Realtime-qPCR assay shown that ART decreased the mRNA levels of proliferation related genes c-Myc, cyclin D1 and PCNA, and reduced the mRNA levels of mesenchymal genes N-cadherin, Vimentin and Snail, but increased the mRNA levels of epithelial marker E-cadherin. Moreover, ART inhibited AKT signaling pathway. In the presence of AKT inhibitor VIII, a pan-AKT inhibitor, ART reduced more cell proliferation, migration and invasion than in the absence of AKT inhibitor VIII, suggesting combination of ART and AKT inhibitor enhanced the anti-cancer effects of ART. Furthermore, the in vivo xenograft tumor model results suggested that ART decreased tumor size and weight, and suppressed AKT signaling. Taken together, our results indicated that ART inhibited ccRCC cell proliferation, colony formation, migration, invasion and tumorigenesis. Combination of ART and AKT inhibitor enhanced the anti-cancer cell proliferation, migration and invasion.

Effect of total glucosides of paeony on the changes of IL-4 and ICAM-1 levels in eczema mouse model serum.

Effect of total glucosides of paeony on the changes of IL-4 and ICAM-1 levels in eczema mouse model serum was investigated. A total of 38 KM mice of SPF grade were divided into 3 groups: the control group (n=10), the model group (n=15) and the treatment group (n=13). The pathological model of chronic eczema in mouse right ear was induced using dinitrochlorobenzene acetone solution. Two ears of mice in the control group and the left ear of mice in the model and treatment groups were smeared with acetone as control. The mice in the treatment group were treated by administration with total glucoside of paeony. The changes of IL-4 and ICAM-1 levels were measured using caudal vein blood collection. The mouse ear weight was measured and the relationship among IL-4 and ICAM-1 levels, ear thickness and treatment time was analyzed. Mouse ear thickness in the model group was higher than that in the treatment and control groups (P<0.05). The weight of the mouse right ear in the model and treatment groups was significantly higher than that of the left ear (P<0.05). Furthermore, The IL-4 and ICAM-1 levels of mice in the model group were higher than that in the treatment and control groups (P<0.05). The IL-4 and ICAM-1 levels of mice in the model and treatment groups increased compared to that before modeling (P<0.05). The IL-4 and ICAM-1 levels of mice were positively correlated with ear thickness in the model group (r=0.865, P=0.002; r=0.833, P=0.009). In addition, the IL-4 level of mice was positively correlated with the ICAM-1 level in the model group (r=0.812, P=0.014). Finally, IL-4 and ICAM-1 may be involved in the pathologic process of chronic eczema. Therefore, the study showed that the total glucosides of paeony may play a role in the treatment of chronic eczema by regulating the IL-4 and ICAM-1 levels.

Long non-coding RNA LINC01410 promotes colon cancer cell proliferation and invasion by inhibiting miR-3128.

A previous study implied that long intergenic non-coding RNA 1410 (LINC01410) promotes angiogenesis and metastasis of gastric cancer. However, the role of LINC01410 in colon cancer (CC) has remained elusive. In the present study, LINC01410 was identified to be highly expressed in CC tissues compared to adjacent normal tissues. It was indicated that high expression of LINC01410 in CC tissues was associated with poor prognosis. Further functional study suggested that LINC01410 knockdown significantly reduced the proliferation and invasive capacity of HT-29 and SW620 cells, and inhibited the cell cycle. Regarding the mechanism, LINC01410 was indicated to serve as a sponge for microRNA (miR)-3128, as evidenced by a luciferase reporter assay. Furthermore, knockdown of LINC01410 significantly increased the levels of miR-3128. In addition, miR-3128 was markedly downregulated in CC tissues compared with that in adjacent normal tissues. A rescue assay revealed that inhibition of miR-3128 significantly abrogated the effects of LINC01410 knockdown on CC cell proliferation and invasion. In conclusion, the present study demonstrated that LINC01410 functions as an oncogene in CC, at least in part by directly inhibiting miR-3128.

Photosynthesis and biochemical responses to elevated O3 in Plantago major and Sonchus oleraceus growing in a lowland habitat of northern China.

A field experiment was carried out to compare the responses to ozone (O3) in two common herbaceous plant species, Plantago major L. and Sonchus oleraceus L., by building open-top growth chambers in situ to simulate O3 stress (+O3, 85±5ppb, 9hr/day for 30days) in a lowland habitat in Inner Mongolia, Northern China. Responses to O3 of gas exchange, chlorophyll a fluorescence, leaf pigment content, antioxidant capability, soluble protein content, membrane lipid peroxidation and dark respiration (Rd) were analyzed. Results showed that elevated O3 exposure significantly reduced the light-saturated net photosynthesis (PNsat), stomatal conductance (gs) and transpiration rate (E) in both species. Although non-significant interactive effect between species and O3 on PNsat was analyzed, the reduction in PNsat in S. oleraceus might be due primarily to the higher fraction of close PSII reaction centers and impaired activities of plant mesophyll cells as evidences by decreased maximum efficiency of PSII photochemistry after dark adapted state (Fv/Fm) and unchanged intercellular CO2 concentration (Ci). Besides, biochemical analysis showed that S. oleraceus had lower antioxidant ability compared to P. major. As a result, S. oleraceus was damaged to the larger extent in terms of lipid peroxidation and visible O3 injury, indicating that S. oleraceus was more sensitive to O3 than P. major. Our results indicated that wild herbaceous plant species growing in a lowland habitat in sandy grassland were sensitive to O3 stress and S. oleraceus can be considered as one of the bio-indicators for high O3 concentration in semi-arid grassland of northern China.

Allocation and simulation study of carbon emission quotas among China's provinces in 2020.

China will form its carbon market in 2017 to focus on the allocation of regional carbon emission quota in order to cope with global warming. The rationality of the regional allocation has become an important consideration for the government in ensuring stable growth in different regions that are experiencing disparity in resource endowment and economic status. Based on constructing the quota allocation indicator system for carbon emission, the emission quota for each province in different scenarios and schemes in 2020 is simulated by the multifactor hybrid weighted Shannon entropy allocation model. The following conclusions are drawn: (1) The top 5 secondary-level indicators that influence provincial quota allocation in weight are as follows: per capita energy consumption, openness, per capita carbon emission, per capita disposable income, and energy intensity. (2) The ratio of carbon emission in 2020 is different from that in 2013 in many scenarios, and the variation is scenario 2 > scenario 1 > scenario 3, with Hubei and Guangdong the provinces with the largest increase and decrease ratios, respectively. (3) In the same scenario, the quota allocation varies in different reduction criteria emphases; if the government emphasizes reduction efficiency, scheme 1 will show obvious adjustment, that is, Hunan, Hubei, Guizhou, and Yunnan will have the largest decrease. The amounts are 4.28, 8.31, 4.04, and 5.97 million tons, respectively.

rhCSF3 accelerates the proliferation of human melanocytes in culture through binding CSF3R and the expression of CSF3R transcripts.

Melanogenic paracrine and autocrine cytokine networks have recently been discovered in vitro between melanocytes and other types of skin cells. Granulocyte colony-stimulating factor receptor (CSF3R) controls the survival, proliferation and differentiation of many kinds of cells, including neutrophils. To understand the function of CSF3R and recombinant human granulocyte colony-stimulating factor (rhCSF3) on melanocyte proliferation, this study compared the expression of CSF3R and the effects of rhCSF3 in primary human melanocytes, neutrophils and HEL 92.1.7 cells. The results show that CSF3R is localized in the cytoplasm and on cell membranes of melanocytes and neutrophils. The percentage of CSF3R(+) melanocytes was higher than CSF3R(+) HEL 92.1.7 cells, but was lower than CSF3R(+) neutrophils. Both CSF3R mRNA and CSF3R protein levels in melanocytes were higher than in HEL 92.1.7 cells, but were lower than in neutrophils. Treatment with rhCSF3 increased the proliferation of human melanocytes, but not their tyrosinase activity. Transcripts of CSF3R in human melanocytes, M14, A375 melanoma and A431 squamous cell carcinoma cells were also detected. Expression of the CSF3R V3 transcript was lower in melanocytes than in M14, A375 melanoma and A431 squamous cell carcinoma cells. In conclusion, rhCSF3 can promote melanocyte proliferation through CSF3R without affecting tyrosinase activity.

The effects of calcipotriol on the dendritic morphology of human melanocytes under oxidative stress and a possible mechanism: is it a mitochondrial protector?

BACKGROUND: Vitiligo is an acquired pigmentary disorder of unknown etiology that is clinically characterized by the development of white macules in the skin related to the selective loss of melanocytes in those areas. Evidence shows that mitochondria might be a unifying target of reactive oxygen species (ROS) generation, cytokine production, catecholamine release and/or alteration of Ca(2+) metabolism that leads to melanocyte loss. OBJECTIVE: To assess the protective effect of calcipotriol on mitochondria of human melanocytes by investigating their dendritic morphology under oxidative stress. METHODS: Human melanocytes were treated with 0.05% H2O2 as well as various concentrations of calcipotriol, after which the retraction velocity of melanocyte dendrites was assessed. Detection of malondialdehyde (MDA) and superoxide dismutase (SOD) was performed as were the mitochondrial membrane potential (MMP) and intracellular calcium concentration ([Ca(2+)]i). Ultrastructural changes of mitochondria in melanocytes were observed by transmission electron microscopy. In addition, the expression of Beclin1, microtubule-associated protein 1 light chain 3 (LC3), dynamin related protein 1 (Drp1), mitofusin 1 (Mfn1) and mitofusin 2 (Mfn2), which are related to autophagy and mitochondrial dynamics, were analyzed by Western blot. RESULTS: Calcipotriol reduced the retraction velocity of melanocyte dendrites. In addition, calcipotriol, from 20nM to 80nM, decreased the level of MDA, increased the activity of SOD, suppressed the reduction of MMP and recovered Ca(2+) homeostasis by reducing [Ca(2+)]i in a concentration-dependent manner. Observation by transmission electron microscopy suggested that calcipotriol might reduce the injury of mitochondria in melanocytes under oxidative stress. Furthermore, the expression of Beclin1, LC3-II/LC3-I, Mfn2 and Drp1 was higher in the calcipotriol-treated melanocytes than in the control or H2O2-treated melanocytes. The level of Mfn1 was almost unchanged, but was higher at a concentration of 80nM calcipotriol than in any other condition. The expression of Mfn2 and Drp1 decreased with increasing calcipotriol concentration. CONCLUSION: Our study demonstrates the antioxidative effect of calcipotriol on melanocytes against oxidative damage. Moreover, calcipotriol could be a promising drug delivery strategy to protect melanocytes against oxidative damage in vitiligo through autophagy or mitophagy.

Activating transcription factor 4 mediates a multidrug resistance phenotype of esophageal squamous cell carcinoma cells through transactivation of STAT3 expression.

Multidrug resistance (MDR) is a major challenge to the clinical treatment of esophageal cancer. The stress response gene activating transcription factor 4 (ATF4) is involved in homeostasis and cellular protection. However, relatively little is known about the expression and function of ATF4 in esophageal squamous cell carcinoma (ESCC) MDR. In this study, we investigate the potential role and mechanisms of ATF4 in ESCC MDR. We demonstrated that overexpression of ATF4 promotes the MDR phenotype in ESCC cells, while depletion of ATF4 in the MDR ESCC cell line induces drug re-sensitization. We also demonstrated that ATF4 transactivates STAT3 expression by directly binding to the signal transducers and activators of transcription 3 (STAT3) promoter, resulting in MDR in ESCC cells. Significantly, inhibition of STAT3 by small interfering RNA (siRNA) or a selective inhibitor (JSI-124) reintroduces therapeutic sensitivity. In addition, increased Bcl-2, survivin, and MRP1 expression levels were observed in ATF4-overexpressing cells. In conclusion, ATF4 may promote MDR in ESCC cells through the up-regulation of STAT3 expression, and thus is an attractive therapeutic target to combat therapeutic resistance in ESCC.

Undifferentiated embryonal sarcoma of the liver mistaken for hepatic abscess in an adult.

Undifferentiated embryonal sarcoma of the liver (UESL) predominantly occurs in children under the age of 10 years, and ~90% of cases occur in children <15 years old. Patients may complain of abdominal pain, fever or other symptoms. No significant decrease has been identified in the hepatic function or elevation of α-fetoprotein, which differentiates UESL from primary carcinomas of the liver. In the present study, a rare and misdiagnosed case of an UESL arising in a male, which was mistaken for a hepatic abscess and retrospectively re-diagnosed, is reported. This case was misdiagnosed as a hepatic abscess initially, and it was diagnosed as UESL subsequent to performing tests, including a type-B ultrasonic scan and computed tomography (CT), and evaluating pathological findings. The rapid recurrence of the tumor in this patient was identified by CT, and this is associated with the malignancy of the disease. Currently, patients with UESL have a poor prognosis as there is not a successful treatment strategy. The present study analyzes the course of diagnosis and potential treatment for the disease.

Preparation of regenerated keratin sponge from waste feathers by a simple method and its potential use for oil adsorption.

In this work, pigeon feathers, a kind of totally waste by-product from the poultry industry, were utilized to fabricate a highly porous keratin sponge in a very simple way by freeze-drying treatment of the dissolved keratin solution, and applied for the first time as an oil adsorbent. An improved method was proposed to dissolve the feather keratin using the inexpensive sodium disulfite as the reducing reagent for sulfitolysis reaction, with a much lower concentration of all involving reaction regents. Moreover, the regenerated keratin sponges obtained a high oil adsorption capacity of above 30 g/g for both liquid paraffin and soybean oil, as well as a good oil holding ability, suggesting that this keratin sponge might be a potential for use as oil adsorbent.

Preparation and characterization of sponge film made from feathers.

Feather wastes generated from poultry farms will pose a problem for disposal, but they are sustainable resources of keratin. Reduction is one of the commonly used methods to obtain soluble keratin from feather. However, the residues generated during feather reduction reaction were rarely investigated. In this study, the residues were transformed into a porous and flexible sponge film by freeze-drying without pretreatment or addition of cross-linking agents. Glycerol was used to alter the physical and chemical characteristics of the sponge film. The film was characterized with a fiber strong stretch instrument, a Fourier transform infrared spectrophotometer, scanning electron microscopy, an elemental analyzer, a differential scanning calorimeter and an automatic air permeability apparatus. Tensile strength and melting point of the sponge film with the optimum glycerol content were 6.2 MPa and 170°C respectively. Due to air permeability of 368 mm/s, the film can potentially be used in medicine, biology, textile, environmental technology, and so on. It is ecologically friendly and will produce additional benefits from the renewable materials. The film was utilized as adsorbents to remove Cr(VI) from aqueous solutions and as a filtering material for air pollution. Its maximum Cr(VI) uptake capacity was about 148.8 mg/g and the removal rate of PM10 was 98.3%.

Rapamycin induces autophagy in the melanoma cell line M14 via regulation of the expression levels of Bcl-2 and Bax.

Cancer therapy with rapamycin has been successfully implemented for kidney cancer, glioblastoma and prostate cancer. However, there are few studies concerning the effects of rapamycin on the treatment of human melanoma. In this study, we investigated whether rapamycin may be a promising strategy for the effective treatment of melanoma and explored the possible mechanism for this by culturing M14 cells in vitro and treating with rapamycin at three concentrations (10, 50 or 100 nmol/l). MDC and LC3B staining, western blot analysis, flow cytometry and transmission electron microscopy were employed. We revealed that rapamycin induced autophagy and inhibited the proliferation of M14 cells in a concentration-dependent manner, Furthermore, western blot analysis revealed an upregulated expression of Bcl-2 and downregulated expression of Bax in M14 cells. In conclusion, rapamycin induced autophagy and inhibited the growth of M14 cells. The mechanism may involve regulation of the expression of Bcl-2 family proteins. Rapamycin appears to be a promising strategy for the effective treatment of melanoma.

Effects of environmental factors on the uptake rates of dissolved nitrogen by a salt-water green alga (Oocystis borgei Snow).

Uptake rates for dissolved nitrogen (DN) by a marine alga (Oocystis borgei) were examined in a (15)N tracer experiment. Maximal uptake rates for all forms of DN were observed at temperatures between 25 and 30°C and at algal concentrations between 3.22 × 10(8) and 4.78 × 10(8 )cell L(-1). Light intensity required to achieve the maximal uptake rate was 45 µmol m(-2) s(-1) for dissolved inorganic nitrogen (DIN = NO(3) (-), NO(2) (-), NH(4) (+)) and methionine, and 126 µmol m(-2) s(-1) for urea. Salinity required to achieve the maximal uptake rate was 12.85 ppt for DIN, 19.89 ppt for urea and 26.2 ppt for methionine.

Composition analysis and application of degradation products of whole feathers through a large scale of fermentation.

PURPOSE: Feathers are one of the most abundant bioresources. They are discarded as waste in most cases and could cause environmental pollution. On the other hand, keratin constituted by amino acids is the main component of feathers. In this article, we reported on biorefined feathers and integrants and application of degraded products. MATERIALS AND METHODS: The fermentation of whole chicken feathers with Stenotrophomonas maltophilia DHHJ in a scale-up of a 5-L bioreactor was investigated in this article. The fermentation process was controlled at 0.08 MPa pressure, 2.5 L/min airflow, and 300 rpm as 100% oxygen saturation level, 40°C, and pH 7.8. RESULTS: Feathers were almost completely degraded in the tested fermentation reaction with the following conditions: 80 g of whole feathers in 3 L fermentation broth for 72 h, seed age of 16 h, 100 mL inoculation amount, and 50% oxygen saturation level. The degraded products contain 397.1 mg/L soluble protein that has mass weight ranging from 10 to 160 kD, 336.9 mg/L amino acids, and many kinds of metal ions. The fermentation broth was evaluated as leaf fertilizer and found to increase plant growth to 82% or 66% for two- or fourfold dilutions, respectively. In addition, in a hair care assay, the broth showed a hair protective function by increasing weight, flexibility, and strength of the treated hair. CONCLUSIONS: The whole feathers were degraded completely by S. maltophilia DHHJ. The degraded product includes many factors to life, such as peptides, amino acids, and mineral elements. It could be applied as leaf fertilizer and hair care product.