[Primary adrenal NK/T cell lymphoma: a clinicopathologic analysis of six cases].

Objective: To investigate the clinicopathologic features of primary adrenal NK/T cell lymphoma (PANKL). Methods: Six cases of PANKL were collected at Henan Provincial People's Hospital from January 2000 to December 2021. The clinicopathologic features including morphology, immunophenotype, treatment and prognosis were retrospectively analyzed, and relevant literature was reviewed. Results: There were two males and four females. The median age was 63 years (ranged from 57 to 68 years). The tumors involved bilateral adrenal glands in 4 cases and unilateral adrenal gland in 2 cases. The main clinical symptom was low back pain without obvious cause. Serum lactate dehydrogenase (LDH) is elevated in five cases. The imaging feature was rapidly enlarging mass initially confined to unilateral/bilateral adrenal glands. Morphologically, the lymphoid cells were mainly medium-sized with a diffuse growth pattern. Coagulative necrosis and nuclear fragmentation were common. Angioinvasion was seen. Immunophenotypically, the neoplastic cells were positive for CD3, CD56 and TIA-1 while CD5 was negative in 5 cases. All cases were positive for EBER by in situ hybridization with more than 80% proliferative activity by Ki-67. Four cases received chemotherapy, one case underwent surgery, and one case underwent surgery with chemotherapy. Follow-up was done in 5 cases; one case was lost to follow-up. Three patients died with a median survival of 11.6 months (3-42 months). Conclusions: PANKL is rare with highly aggressive clinical presentation and poor prognosis. Accurate diagnosis entails correlation of histomorphology, immunohistochemistry, EBER in situ hybridization and clinical history.

[Metal artifact reduction and clinical verification in oral and maxillofacial region based on deep learning].

Objective: To construct a kind of neural network for eliminating the metal artifacts in CT images by training the generative adversarial networks (GAN) model, so as to provide reference for clinical practice. Methods: The CT data of patients treated in the Department of Radiology, West China Hospital of Stomatology, Sichuan University from January 2017 to June 2022 were collected. A total of 1 000 cases of artifact-free CT data and 620 cases of metal artifact CT data were obtained, including 5 types of metal restorative materials, namely, fillings, crowns, titanium plates and screws, orthodontic brackets and metal foreign bodies. Four hundred metal artifact CT data and 1 000 artifact-free CT data were utilized for simulation synthesis, and 1 000 pairs of simulated artifacts and metal images and simulated metal images (200 pairs of each type) were constructed. Under the condition that the data of the five metal artifacts were equal, the entire data set was randomly (computer random) divided into a training set (800 pairs) and a test set (200 pairs). The former was used to train the GAN model, and the latter was used to evaluate the performance of the GAN model. The test set was evaluated quantitatively and the quantitative indexes were root-mean-square error (RMSE) and structural similarity index measure (SSIM). The trained GAN model was employed to eliminate the metal artifacts from the CT data of the remaining 220 clinical cases of metal artifact CT data, and the elimination results were evaluated by two senior attending doctors using the modified LiKert scale. Results: The RMSE values for artifact elimination of fillings, crowns, titanium plates and screws, orthodontic brackets and metal foreign bodies in test set were 0.018±0.004, 0.023±0.007, 0.015±0.003, 0.019±0.004, 0.024±0.008, respectively (F=1.29, P=0.274). The SSIM values were 0.963±0.023, 0.961±0.023, 0.965±0.013, 0.958±0.022, 0.957±0.026, respectively (F=2.22, P=0.069). The intra-group correlation coefficient of 2 evaluators was 0.972. For 220 clinical cases, the overall score of the modified LiKert scale was (3.73±1.13), indicating a satisfactory performance. The scores of modified LiKert scale for fillings, crowns, titanium plates and screws, orthodontic brackets and metal foreign bodies were (3.68±1.13), (3.67±1.16), (3.97±1.03), (3.83±1.14), (3.33±1.12), respectively (F=1.44, P=0.145). Conclusions: The metal artifact reduction GAN model constructed in this study can effectively remove the interference of metal artifacts and improve the image quality.

[Establishment and preliminary application of organoids in ovarian cancer].

Objective: To explore the establishment and application of ovarian cancer organoids. Methods: Fresh ovarian tumor tissues, obtaining from patients underwent surgery in the First Affiliated Hospital of Nanjing Medical University between October 2021 and March 2022, were collected, enzymatic degraded, digested, and embedded into matrigel to establish organoids. A total of 32 ovarian cancer samples were collected. Hematoxylin eosin (HE) staining and immunofluorescence (IF) procedure were used to verify the morphological structure of organoids and their expression of molecular markers. 3D cyto-live or dead assay was used to detecte the live or dead cells in organoids. Carboplatin with a concentration ranging from 5 to 80 µmol/L (5, 10, 20, 40, 80 µmol/L) was added to organoids to calculate the 50% inhibitory concentration (IC50) in different organoids. Results: (1) Organoids from a total of 32 patients were established, of which 18 cases could be passaged stably in the long term in vitro, while 14 could be passaged in the short time. The average amplification time of long-term passage in vitro was over 3 months, and the longest reached 9 months. (2) In HE staining, significant nuclei atypia and local micropapillary structures were observed in organoids. IF staining revealed that ovarian cancer organoids expressed molecular markers similar to primary tumor tissues, such as Pan cytokeratin (Pan-CK), p53, paired box gene 8 (PAX8), and Wilms tumor gene 1 (WT1). (3) In 3D cyto-live or dead assay, a large number of apoptotic cells were observed inside and around the organoids after added carboplatin. The sensitivity to carboplatin varied in 18 organoids could amplify in the long term, with an average IC50 of (29.5±15.8) µmol/L. Moreover, IC50 values of 4 organoids derived from patients received neoadjuvant chemotherapy were much higher than the 14 organoids which did not received neoadjuvant chemotherapy [(48.7±11.3) µmol/L vs (24.0±12.1) µmol/L; t=3.429, P=0.022]. Conclusions: Organoids recapitulate ovarian cancers in vitro and could be stably passaged. Organoids derived from patients received neoadjuvant chemotherapy have higher resistance to carboplatin.

[Relationship between parental reproductive age and the risk of overweight and obesity in offspring].

Objective: To explore the role of parental reproductive age on the risk of overweight and obesity in offspring. Methods: The participants were derived from physical examination data of students aged 6-18 years in seven provinces in China, and questionnaire survey was used to collect demographic characteristics and lifestyle information of the students and their parents. A total of 41 567 children with complete data were included. According to the restricted cubic spline curve, maternal reproductive age was divided into three categories, 14-23, 24-28, and 29-38 years, and paternal reproductive age was divided into 14-23, 24-30, and 31-42 years. Multivariate logistic regression model was used to analyze the association between parental reproductive age and parental nutritional status and the risk of overweight and obesity in offspring. Results: The mean age of 41 567 children was (10.6±3.2) years, and the mean paternal and maternal age were (27.9±4.4) years and (25.8±4.0) years, respectively. The detection rate of overweight and obesity was 23.4%. After adjusting factors of children diet and behaviors, the OR(95%CI)of offspring overweight and obesity in groups of fathers aged 24-30 years and mothers aged 24-28 years was 1.11 (1.04-1.18) and 1.16 (1.08-1.24), respectively. When none parents were overweight and obese, the difference of obesity risk was not statistically significant. When both parents were overweight and obese, the OR(95%CI)of offspring overweight and obesity in groups of fathers aged 24-30 years and mothers aged 14-28 years old was 1.27 (1.00-1.62) and 1.33 (1.07-1.65) respectively. Conclusion: Parental reproductive age and parental overweight and obesity status may both increase the risk of overweight and obesity in offspring, with a significant interaction effect.

[Spindle cell type squamous dysplasia of the esophagus: a clinicopathological analysis].

Objective: To investigate the clinicopathological features and significance of spindle cell type squamous dysplasia of the esophagus. Methods: The clinicopathological data of 37 cases of spindle cell type squamous dysplasia of esophagus were collected retrospectively at People's Liberation Army Joint Logistics Support Force 989 Hospital (formerly 152 Hospital), Pingdingshan, China, from 2009 to 2019. The histological and immunohistochemical characteristics were analyzed, with a literature review. Results: The median age of the 37 patients was 65 years (range 47-81 years), while the ratio of men to women was 1.5∶1.0. There were 4 cases in the upper esophagus, 31 in the middle esophagus and 2 in the lower esophagus. The median diameter of the lesions was 14 mm (range 3-40 mm). According to the Paris classification, 11 cases were 0-Ⅱa, 14 cases were 0-Ⅱb, 3 cases were 0-Ⅱb and 0-Ⅱa, and 9 cases were 0-Ⅱc. Under endoscope, the lesional mucosa was reddish. The micro-vessels were dilated, with various shapes and density. Histologically, tumor cells and nuclei were spindle shaped or elongated spindle shaped, with considerable homogeneity, dark nuclei and delicate or slightly thickened chromatin. The mitosis was conspicuous, and atypic mitoses were seen; the cytoplasm was acidophilic, and the intercellular bridge was obvious. The cells were dense and often lost polarity, but still arranged in parallel, mostly perpendicular to the basement membrane. Spindle cells often involved the whole layer of epithelium, with no gradient maturation and differentiation of normal squamous epithelium. The tumor was well demarcated. The spindle cells often invaded lamina propria. There were 15 cases with focal high-grade dysplasia and superficial invasive squamous cell carcinoma. Immunohistochemical staining showed that the mutation rate of p53 was 41.4% (12/29), the median of Ki-67 labeling index was 40% (range 20%-80%), and the abnormal distribution pattern of Ki-67 was 29 (100%). According to the initial pathological diagnosis, there were 6 cases of low-grade dysplasia, 4 cases of atypical epithelial cells and 27 cases of high-grade dysplasia and superficial invasive squamous cell carcinoma. Conclusions: Spindle tumor cells have moderate to severe atypia, and some tumors show invasive pattern. P53 mutation and Ki-67 abnormal distribution pattern indicate that they are high-grade dysplasia of esophageal squamous epithelium. The unique characteristics of spindle tumor cells suggest that they may represent a spindle cell subtype in the morphological spectrum of esophageal squamous dysplasia. When the knowledge of the lesion is insufficient, it can be easily misdiagnosed or missed.

[Clinical pathological features of 180 cases with primary esophageal malignant melanoma].

Objective: To investigate the clinical pathological and epidemiological characteristics of primary esophageal malignant melanoma (PMME). Methods: The clinical pathology data of 180 PMME patients in the esophageal cancer database of the key laboratory of esophageal cancer research in Henan Province from 1973 to 2016 were collected, of which 136 were male, aged (58.5±9.0) years, 44 were female, aged (56.7±12.2) years. Kaplan-Meier and Log rank test were used for survival analysis, Cox regression scale model was used for risk factor analysis. Results: The incidence of PMME is 0.036% (180/500, 000), mostly were male (about 3∶1 for men: female). The common sites of PMME were the lower part of the esophagus (48.9%, 85/174), followed by the middle section of the esophagus (46.0%, 80/174) and the upper part of the esophagus (5.2%, 9/174). No black particles were seen in the PMME cells of 3 patients under microscope, and strong positive expressions of Melan-A and HMB453 were observed in these 3 patients by immunohistochemical results. Of the 129 patients who had a routine preoperative esophageal biopsy, 69 were undiagnosed with PMME (53.5%). The medium survival time of the whole group was 7.9 months, and the survival rates of 1, 2, 3, 5 years were 25.0%, 7.9%, 6.6% and 1.3%, respectively. The univariate analysis showed that N, M, TNM phase and radiotherapy were related to the overall survival of patients (P<0.05). Multivariate analysis showed that TNM phase and radiotherapy were the independent risk factors for overall survival of patients (P<0.05). Conclusions: PMME is more common in men, the common site of the disease is the lower part of the esophagus. The preoperatively missed diagnosis rate of Chinese PMME is high. TNM phase and radiotherapy are the independent risk factors for overall survival of patients.

[Clinicopathological features of basal cell type dysplasia of esophagus].

Objective: To investigate the clinicpathological features of basal cell type dysplasia of the esophagus. Methods: The clinicopathological data of 71 cases of basal cell type dysplasia of esophagus were collected at the People's Liberation Army Joint Logistics Support Force 989 Hospital, from 2009 to 2019, and the histomorphologic characteristics and immunophenotype were evaluated. The relevant literature was reviewed. Results: The ratio of male to female patients was 1.6∶1.0, and the median age was 65 years (range 48-81 years). The tumors were located in the upper segment of the esophagus in four cases (5.6%), the middle segment in 54 cases (76.1%), and the lower segment in 13 cases (18.3%).The median maximal tumor diameter was 12.0 mm (range 3-42 mm). According to Paris Classification, 0-Ⅱb accounted for 42.3% (30/71) of the cases. Under endoscope, the lesions were reddish with abnormal mucosal microvessels. Histologically, the neoplastic cells were small, with a high nuclear-cytoplasmic ratio, similar to basal cells, and uniform in morphology. The structural atypia was characterized by dense and disordered tumor cells, loss of basal cell polarity, and absence of normal squamous differentiation gradient. In 10 cases, the tumors were confined to the lower part of the epithelium. The tumor cells were smaller and more uniform in shape, and extend to the superficial lamina propria. Sixty-one tumors involved at least the entire layer of the upper cortex. There were 31 cases of neoplasms with superficial invasive carcinoma. The types of neoplasms included typical squamous cell carcinoma, basaloid squamous cell carcinoma, small cell neuroendocrine carcinoma, squamous cell carcinoma with sebaceous adenoid carcinoma, and differentiation of glandular/ductal epithelioid carcinoma. Immunohistochemical staining showed that the mutant expression rate of p53 protein was 41.5% (17/41). All 41 cases (100.0%) showed abnormal distribution pattern of Ki-67. According to the initial pathologic diagnosis, there were 18 cases of low grade dysplasia, 12 cases of atypical epithelial cells, and 41 cases of high grade dysplasia and superficially invasive carcinoma. Conclusions: Basal cell type dysplasia has unique morphologic characteristics and represents a tumor subtype in the morphologic lineage of esophageal squamous dysplasia. Tumor cells of basal cell type dysplasia, especially those distributed only in the lower part of the stratified squamous epithelium, may be tumor stem cells at the earliest stage of esophageal carcinogenesis and have multidirectional differentiation potential. When the tumor is confined to the lower part of the stratified squamous epithelium, it does not meet the diagnostic criteria for esophageal squamous dysplasia as defined by the current WHO classification.

The relationship between endoplasmic reticulum stress and liver function, insulin resistance and vascular endothelial function in patients with non-alcoholic fatty liver disease.

OBJECTIVE: The aim of the study was to investigate the relationship between ER stress and liver function, insulin resistance and vascular endothelial function in patients with non-alcoholic fatty liver disease. PATIENTS AND METHODS: A total of 95 patients with non-alcoholic fatty liver disease were selected. They were admitted to our hospital from November 2016 to January 2019. A total of 90 cases of obese patients without fatty liver were selected as control group during the same period. The levels of ER stress marker protein were compared between the two groups, and the relationship between ER stress and liver function, insulin resistance, and vascular endothelial function was analyzed. RESULTS: The protein level of ER stress markers in the test group was significantly higher than that in the control group (p<0.05). The liver function index and insulin resistance level were significantly higher than those in the control group (p<0.05). The level of vascular endothelial function was significantly lower than that of the control group (p<0.05). Pearson correlation analysis showed that ER stress marker protein was positively correlated with liver function and insulin resistance (p<0.05), while ER marker protein was negatively correlated with vascular endothelial function (p<0.05). CONCLUSIONS: Liver function and insulin resistance are closely related to ER stress in patients with non-alcoholic fatty liver disease. Insulin resistance is one of the factors inducing and aggravating endothelial dysfunction.

[Effects of fecal microbiota transplantation in different routes on the clinical efficacy of slow transit constipation].

Objective: To evaluate the efficacy and safety of the fecal microbiota transplantation (FMT) in the different route administration for slow transit constipation (STC). Methods: A retrospective cohort study was conducted. The clinical data of 270 STC patients who voluntarily received FMT treatment in the Tenth People's Hospital of Tongji University from May 2018 to May 2019 were collected. Non-relative healthy adult standard donors were applied. The treatment routes of bacterial flora transplantation included nasojejunal tube (nasal enteral tube group, 120 cases), oral enterobacterial capsule treatment (oral capsule group, 120 cases), and colonoscopy infusion (colonoscopy group, 30 cases). The efficacy and safety of treatment among the three groups were compared. Results: Transplanted bacteria of three groups were extracted from 100 g of fresh feces. All the patients successfully completed the transplantation. The waiting time for the nasal enteral tube group, oral capsule group and colonoscopy group was (1.5±0.5) d, (0.4±0.3) d and (3.6±0.8) d respectively; the cost of establishing the transplantation path was (495±20) yuan, (25±10) yuan and (1420±45) yuan respectively, whose differences were statistically significant (F=9.210, P=0.03; F=10.600,P=0.01). The clinical improvement rates at 1 month after FMT treatment in the nasojejunal tube group, oral capsule group and colonoscopy group were 74.2% (89/120), 60.0% (72/120) and 53.3% (16/30) respectively, whose difference was statistically significant (χ(2)=5.990, P<0.05). The clinical improvement rates at 3 months after treatment were 71.1% (69/97), 53.6% (45/84), and 44.0% (11/25) respectively, whose difference was statistically significant (χ(2)=7.620, P<0.05). The incidence of adverse reactions in the colonoscopy group was 76.7% (23/30), which was higher than that in the nasal nasojejunal group (39.2%, 47/120) and oral capsule group (21.7%, 26/120). The most common adverse reactions in the nasojejunal tube group, oral capsule group and colonoscopy group were respiratory discomfort (17.5%, 21/120), nausea and vomiting (10.0%, 12/120), and diarrhea (36.7%, 11/30). During the 3-month follow-up after treatment, no FMT-related adverse reactions were reported. Conclusions: The nasojejunal tube route has stable clinical efficacy and operability, while the oral capsule route has shorter waiting time and less cost. However, the adverse reactions caused by different transplantation methods are different, thus personalized transplantation method should be recommended.

[Effects of copper and cadmium ions on protective enzyme activity in Oncomelania hupensis].

OBJECTIVE: To evaluate the effects of Cu2+ and Cd2+ at different concentrations on superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) activity in Oncomelania hupensis. METHODS: Cu2+- and Cd2+-containing solutions were prepared at 7 concentrations, and O. hupensis snails were exposed to the solutions for 24 h, of 15 snails in each concentration. Then, the snail body was collected following removal of the snail shell and homogenated, and the SOD, CAT and POS activities were detected in the supernatants. RESULTS: With the increase of the Cu2+ concentration, the SOD activity appeared a rise followed by a reduction in O. hupensis snails, and the CTA activity appeared a decline-rise-decline tendency, while the POD activity showed a tendency towards rise followed by decline. With the increase of the Cd2+ concentration, the SOD activity appeared a rise followed by a reduction in O. hupensis snails, and the CTA activity appeared a decline- rise- decline tendency, while the POD activity showed a decline-rise-decline tendency. CONCLUSIONS: Exposure to Cu2+ and Cd2+ at high concentrations results in a decline in the activity of SOD, CAT and POD in O. hupensis at the same time.

[Acute toxicity of four heavy metal ions to Oncomelania hupensis].

OBJECTIVE: To assess the acute toxicity of Cu2+, Cd2+, Hg2+ and Pb2+ to Oncomelania hupensis. METHODS: Cu2+, Cd2+, Hg2+ and Pb2+ solutions were prepared at five concentrations, and 10 snails were exposed to each concentration for 24, 48, 72 h and 96 h. Then, the inhibition of snail activity and snail death was observed, and the half maximal effective concentration (EC50) and median lethal concentrations (LC50) were estimated. RESULTS: The 24, 48, 72 h and 96 h EC50 values of Cu2+, Cd2+, Hg2+ and Pb2+ were 0.74, 0.56, 0.46, 0.37 mg/L, 4.79, 3.52, 1.70, 1.26 mg/L, 1.90, 1.49, 0.83, 0.76 mg/L and 21.40, 9.98, 7.90, 5.42 mg/L for snails, respectively. The 96 h LC50 values of Cu2+, Cd2+, Hg2+ and Pb2+ were 0.43, 2.96, 1.12 mg/L and 12.22 mg/L for snails, the safe concentrations were 0.004 3, 0.029 6, 0.011 2, 0.122 2 mg/L, respectively. CONCLUSIONS: Cu2+ shows a high acute toxicity to snails, and Cd2+ and Hg2+ exhibit a moderate acute toxicity to snails, while Pb2+ is lowly toxic to snails.

MiR-296-5p suppresses papillary thyroid carcinoma cell growth via targeting PLK1.

OBJECTIVE: Our study aimed to explore the effects of miRNA-296-5p on the biological behaviors of papillary thyroid carcinoma (PTC) cells and its potential mechanism. PATIENTS AND METHODS: Twenty-eight PTC tissues and the corresponding non-cancerous tissues were collected. Real Time-quantitative Polymerase Chain Reaction (RT-qPCR) analysis was performed to detect the expression levels of miR-296-5p in PTC tissues and the adjacent non-cancerous tissues. Besides, the different endogenous expression levels of miR-296-5p in PTC cell line (K1) and normal thyroid gland cell line (Nthy-ori3-1) were also detected by RT-qPCR. Bioinformatics analysis, Western blot and Dual-Luciferase reporter gene assay were performed to demonstrate whether polo-like kinase 1 (PLK-1) was a downstream target of miR-296-5p. Subsequently, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay, flow cytometry analysis, colony formation assay and TUNEL assay were performed to estimate whether PLK1 down-regulation could attenuate the malignant behaviors of PTC cells in vitro. RESULTS: RT-qPCR results showed that the expression level of miR-296-5p was significantly down-regulated in PTC tissues and cells, indicating that miR-296-5p may participate in PTC development. We predicted target genes of miR-296-5p by bioinformatics and identified PLK1 as a target gene of miR-296-5p. By Western blot and Dual-Luciferase reporter gene assay, we confirmed that miR-296-5p was partially complement to PLKl mRNA 3'UTR sequence and inhibited PLK1 expression at the post-transcriptional level. In vitro experiments suggested that the transfection of miR-296-5p mimics into K1 cells suppressed cell proliferation, inhibited cell clone formation, arrest the cell cycle in G2/M phase and induced apoptosis. Importantly, PLK1 reversed the inhibitory effects of miR-296-5p on biological behaviors of PTC. CONCLUSIONS: MiR-296-5p influences the biological behaviors of PTC by regulating PLK1. These findings provide a new perspective for the molecular mechanism of PTC pathogenesis and also contribute to developing new targets and methods for PTC treatment.

Determination of Place of Residence Using the Gene Information of Plants Carried by the Human Body.

ABSTRACT: Objective To identify the plant species using the DNA sequence of plant pollen from lung tissues of a unidentified body, infer the possible long-term places of residence of the deceased according to the distribution area of the pollen in the lung tissues, therefore narrow the scope of criminal investigation and provide clues for case solving. Methods Lung tissues were extracted from the deceased, total DNA was extracted by the mCTAB method. Gene fragments of the two plant DNA barcodes, matK and rbcL, were acquired using specific primers for amplification, then sequenced. The DNA sequences of target gene fragments were acquired through bioinformatics analysis. The sequences were combined with reference sequence data. Phylogenetic analysis was made to identify the species that the DNA sequences belonged to. The places where the deceased could have lived for a long time were inferred, according to the distribution information of plant species. Results Gene fragments of 32 plant species which belonged to 31 genera of 27 families were in the lung tissues of the deceased. Among them, plants of 9 genera that had certain indicative function were mainly endemic plants from Hainan, Guangdong, Guangxi and Yunnan. These results showed that the deceased may have stayed in these areas for a long time before death. After further investigation, the victim was confirmed to have come from a county in southern Guangxi, which was in accordance with the research results. Conclusion The method of using gene information of plants from lung tissues of human bodies to infer places of residence can assist inference of the places where the deceased could have lived for a long time. The present study may also provide new ideas for locating sources of the corpses in cases with unidentified victims.

MicroRNA-615-3p promotes the osteoarthritis progression by inhibiting chondrogenic differentiation of bone marrow mesenchymal stem cells.

OBJECTIVE: To investigate whether microRNA-615-3p participates in the development and progression of osteoarthritis by regulating chondrogenic differentiation of bone marrow mesenchymal stem cells. MATERIALS AND METHODS: Bone marrow mesenchymal stem cells (BMSCs) were isolated from rat bone marrow and identified by flow cytometry. After chondrogenic differentiation was induced in BMSCs, expression levels of chondrogenic-specific genes were then detected by quantitate Real-time polymerase chain reaction (qRT-PCR). Expression levels of inflammatory cytokines were detected by enzyme-linked immunosorbent assay (ELISA). Protein expression of SOX9 after overexpression or knockdown of microRNA-615-3p was detected by Western blot, respectively. RESULTS: MicroRNA-615-3p was down-regulated in the process of chondrogenic differentiation of BMSCs. The mRNA expressions of chondrogenic-specific markers, COL2A1, COL10A1, ACAN and MATN3 were decreased after microRNA-615-3p overexpression in BMSCs. Overexpressed microRNA-615-3p down-regulated protein expression of SOX9. Expression levels of inflammatory cytokines, including interleukin-1 (IL-1), interleukin-6 (IL-6), interleukin-α (IL-α) were increased after overexpression of microRNA-615-3p, while inhibition of microRNA-615-3p expression obtained the opposite result. In addition, overexpression of SOX9 rescued the effect induced by microRNA-615-3p on inflammatory cytokines. CONCLUSIONS: MicroRNA-615-3p participates in the development and progression of osteoarthritis by increasing the expressions of inflammatory cytokines and inhibiting chondrogenic differentiation of BMSCs.

Diagnostic accuracy of natriuretic peptides for acute heart failure: a review.

The rising incidence of and the cost associated with heart failure have made it increasingly imperative to accurately diagnose heart failure upon presentation. Correctly identifying heart failure in an Emergency Department is extremely challenging, and according to estimates, is only confirmatory in approximately 40-50% of patients. For an accurate diagnosis of heart failure and the consequent treatment, there needs to be more accurate test relying on biochemical factors as opposed to general symptoms that patients are experiencing. Natriuretic peptides are now utilized in routine tests for heart disease diagnosis in emergency departments as it is relatively low cost, easy to use and is a quick way to exclude heart failure as a reason for dyspnea. In this review, we detail the role and value of individual natriuretic peptides, particularly BNP, NT-proBNP, and MR-proANP, in diagnosing acute heart failure.

Global land-water nexus: Agricultural land and freshwater use embodied in worldwide supply chains.

As agricultural land and freshwater inextricably interrelate and interact with each other, the conventional water and land policy in "silos" should give way to nexus thinking when formulating the land and water management strategies. This study constructs a systems multi-regional input-output (MRIO) model to expound global land-water nexus by simultaneously tracking agricultural land and freshwater use flows along the global supply chains. Furthermore, land productivity and irrigation water requirements of 160 crops in different regions are investigated to reflect the land-water linkage. Results show that developed economies (e.g., USA and Japan) and major large developing economies (e.g., mainland China and India) are the overriding drivers of agricultural land and freshwater use globally. In general, significant net transfers of these two resources are identified from resource-rich and less-developed economies to resource-poor and more-developed economies. For some crops, blue water productivity is inversely related to land productivity, indicating that irrigation water consumption is sometimes at odds with land use. The results could stimulus international cooperation for sustainable land and freshwater management targeting on original suppliers and final consumers along the global supply chains. Moreover, crop-specific land-water linkage could provide insights for trade-off decisions on minimizing the environmental impacts on local land and water resources.

Optimization of culture conditions for penicilazaphilone C production by a marine-derived fungus Penicillium sclerotiorum M-22.

The aim of this study was to optimize the culture conditions of a marine-derived fungus Penicillium sclerotiorum M-22 for the production of penicilazaphilone C (PAC), a novel azaphilonidal derivative exhibiting broad cytotoxic and antibacterial effects. By single factor experiments, the effects to the production of PAC of aged seawater concentration, initial pH values, fermentation time, carbon sources, nitrogen sources and inorganic salt sources were investigated individually. Response surface methodology (RSM) analysis was adopted to investigate the interactions between variables and determine the optimal values for maximum PAC production. Evaluation of the experimental results signified that the optimum conditions for maximum production of PAC (19·85 mg l-1 ) in 250 ml Erlenmeyer flask were fermentation time 24·83 days, pH of 7·00, corn meal concentration of 10·72 g l-1 , yeast extract concentration of 4·58 g l-1 , crude sea salt concentration of 20·59 g l-1 . Production under optimized conditions increased to 1·344-fold comparing to its production prior to optimization. The higher PAC production and the penicilazaphilone C -producing marine fungus would be provide a promising alterative approach for industrial and commercial applications. SIGNIFICANCE AND IMPACT OF THE STUDY: Penicilazaphilone C (PAC) was a novel azaphilonidal derivative which had exhibited selective cytotoxicity and antibacterial activity. To further enhance production of PAC by optimizing fermentation conditions of Penicillium sclerotiorum M-22 would provide a promising alterative approach for industrial and commercial applications. We used the single factor test to determine the key factors which influence the PAC production. Then through the Response surface methodology and Box-Behnken design to determine the best fermentation condition for maximum production of PAC. Through these experimental designs and analysis will help us improve experimental efficiency and save time and materials.

[High expression of hnRNPAB/Kap1 together promote poor prognosis in HCC].

Objective: To further understand the interaction protein spectrum of heterogeneous ribonucleoprotein AB (hnRNP AB), and to investigate their clinical significance in hepatocellular carcinoma (HCC). Methods: We carried out mass spectrometry to reveal the specific peptides of KRAB-associated protein 1 (Kap1) and hnRNPAB, and verified their interaction by immunocoprecipitation and western blotting. Expression of hnRNPAB/Kap1 proteins were detected by immunohistochemical staining in the tissue microarrays. Categorical data were analyzed by the chi square test or Fisher exact test; enumeration data between groups were compared using Student t-test or Wilcocon signed rank test; the cumulative recurrence and survival rates were evaluated using the Kaplan-Meier method and the differences were assessed using the log-rank test. Results: We identified Kap1 as a molecular partner for hnRNPAB in HCCLM3 cells and HepG2 cells as well. We found that the 5-year survival rate of the Kap1high patients was significantly lower than the survival rate of those of the Kap1low group (36% vs 59% , HR = 1.67, P < 0.001). Similarly, Kap1high HCC patients had the poorest prognosis at 5-years, with higher cumulative recurrence rate than Kap1low patients (72% vs 54%, HR = 1.66, P = 0.001). Univariate and Multivariate analyses revealed that hnRNPAB /Kap1 alone (HR = 1.35 /1.28, P = 0.001) or in combination with Kap1 (HR =1.24 /1.27, P < 0.05) were independent prognostic indicators for overall survival and time to recurrence. Conclusion: In HCC cells, hnRNPAB and Kap1 form protein complexes. The expression levels of hnRNPAB alone or in combination with Kap1 in HCC patients are important because they provide not only a predictor for HCC prognosis but also a therapeutic target for future studies.