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Introduction: Despite rapid advances in liquid biopsy for circulating tumor DNA (ctDNA), its prognostic value for venous thromboembolism (VTE) in patients with cancer is underexplored, particularly in underserved and minoritized populations., Methods: We analyzed data from 1,038 cancer patients who underwent ctDNA measurement for oncologic care at a large safety-net hospital system in the US. We investigated the association between ctDNA and VTE after adjusting for cancer type, stage, treatment, and time from initial diagnosis using Fine-Gray models. We further assessed the discrimination of the genetic, clinical-only, and combined models using the area under the time-dependent receiver operating characteristic curve (AUC)., Results: The presence of pathogenic ctDNA was independently associated with VTE after adjusting for clinical variables. Independent of tumor type, the number of pathogenic ctDNA mutations was predictive of future VTE risk (adjusted subdistribution hazard ratio 2.75, 1.94, and 1.38 for ≥3, 2, and 1 pathogenic mutation, respectively, compared to none; p<0.0001). The association was primarily driven by mutations in KRAS, PTEN, CDKN2A, NF1, and EGFR genes. Compared to the clinical-only model (AUC 0.71, 95% CI 0.64-0.76), the combined clinical and ctDNA model had a numerically higher time-dependent AUC (AUC 0.74, 95% CI 0.67-0.80)., Conclusions: CtDNA testing may serve as an adjunctive tool to clinical risk assessment models in cancer patients to improve personalized VTE risk assessment and management., (Copyright © 2024 International Society on Thrombosis and Haemostasis. All rights reserved.)

Background: There is uncertainty in the management of cancer-associated isolated splanchnic vein thrombosis (SpVT)., Objectives: To describe the natural history of SpVT by cancer type and thrombus composition and to review anticoagulation (AC) practices and associated rates of usual-site venous thromboembolism (VTE), major and clinically relevant nonmajor bleeding (MB/CRNMB), recanalization/progression, and mortality., Methods: We performed a retrospective cohort study in patients with SpVT at 2 cancer care centers in Houston, Texas. We estimated the incidence of usual-site VTE and MB/CRNMB at 6 months using competing risk methods and examined venous patency in a subset of patients with repeat imaging. We assessed associations with mortality using Cox regression., Results: Among 15 342 patients with an incident cancer diagnosis from 2011 to 2020, we identified 298 with isolated SpVT. Patients with hepatocellular carcinoma (HCC) and SpVT (n = 146) had the highest disease prevalence (20%), lowest rate of AC treatment (2%), and similar rate of usual-site VTE (4.2%) vs those without SpVT (5.2%) at 6 months, though tumor thrombus vs bland was associated with worse overall survival. In patients with non-HCC bland SpVT (n = 114), AC (n = 37) was more common in those with non-upper gastrointestinal cancers and fewer comorbidities. AC was associated with more recanalization (44% vs 15%, P = .041) but no differences in usual-site VTE, MB/CRNMB, or mortality at 6 months., Conclusion: Cancer-associated isolated SpVT is a common but heterogeneous thrombotic disease that is treated differently from usual-site VTE. Tumor thrombus is a negative prognostic factor. Initiation of AC in bland thrombi requires judicious consideration of thrombotic and bleeding risk., Competing Interests: Declaration of competing interests The authors declare no competing financial interests., (Copyright © 2024 International Society on Thrombosis and Haemostasis. Published by Elsevier Inc. All rights reserved.)

Background: Pancreatic cancer is a common malignancy with poor prognosis and limited treatment. Here we aimed to investigate the role of host chromosomal instability (CIN) and tumor microbiome in the prognosis of pancreatic cancer patients., Methods: One hundred formalin-fixed paraffin-embedded (FFPE) pancreatic cancer samples were collected. DNA extracted from FFPE samples were analyzed by low-coverage whole-genome sequencing (WGS) via a customized bioinformatics workflow named ultrasensitive chromosomal aneuploidy detector., Results: Samples are tested according to the procedure of ultrasensitive chromosomal aneuploidy detector (UCAD). We excluded 2 samples with failed quality control, 1 patient lost to follow-up and 6 dead in the perioperative period. The final 91 patients were admitted for the following analyses. Thirteen (14.3%) patients with higher CIN score had worse overall survival (OS) than those with lower CIN score. The top 20 microbes in pancreatic cancer samples included 15 species of bacteria and 5 species of viruses. Patients with high human herpesvirus (HHV)-7 and HHV-5 DNA reads exhibited worse OS. Furthermore, we classified 91 patients into 3 subtypes. Patients with higher CIN score (n =13) had the worst prognosis (median OS 6.9 mon); patients with lower CIN score but with HHV-7/5 DNA load (n = 24) had worse prognosis (median OS 10.6 mon); while patients with lower CIN score and HHV-7/5 DNA negative (n = 54) had the best prognosis (median OS 21.1 mon)., Conclusions: High CIN and HHV-7/5 DNA load were associated with worse survival of pancreatic cancer. The novel molecular subtypes of pancreatic cancer based on CIN and microbiome had prognostic value., Competing Interests: Competing interest No benefits in any form have been received or will be received from a commercial party related directly or indirectly to the subject of this article., (Copyright © 2024 First Affiliated Hospital, Zhejiang University School of Medicine in China. Published by Elsevier B.V. All rights reserved.)

Carboxymethyl chitosan and resistant starch exhibit good performance in diabetes regulation. We prepared carboxymethyl chitosan - resistant starch complex. Test the properties of composite resistant starch by using X-ray diffraction, water contact angle, infrared spectroscopy, and scanning electron microscopy, interactions with intestinal microbiota and mouse experiments were also conducted. The results indicated that the composite resistant starch had a good effect on promoting the proliferation of probiotics on Bifidobacterium and a significant inhibitory effect on Escherichia coli than resistant starch (P < 0.05). After administration, the water intake and weight of diabetic mice were significantly reduced. The blood glucose of diabetic mice was also reduced, and oral glucose tolerance showed that the glucose degradation rates of composite resistant starch were significantly improved compared to model mice. Cholesterol, triglycerides, high-density lipoprotein and low-density lipoprotein were significantly lower than those in the diabetes group (P < 0.05). The diversity of the gut microbiota was also proven., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Cadmium (Cd) is a common contaminant in aquatic environments. However, little is known about the mechanisms underlying Cd toxicity in the freshwater snail Cipangopaludina cathayensis (Heude, 1890). This study to investigate the toxic effects of Cd on the standard metabolism, antioxidant activities, immune function, and hepatopancreas transcriptome profiles of C. cathayensis. C. cathayensis was exposed to 0.25, 0.5, 1.0, or 1.5 mg/L Cd for 3 h, with results showing that Cd significantly inhibited oxygen consumption and ammonia excretion and disrupted the respiratory metabolism of C. cathayensis. In addition, the O:N ratio dropped below 7, indicating that C. cathayensis may rely exclusively on proteins as an energy source under Cd stress. To understand how Cd impacts the antioxidant activities, immune function, and transcriptional profiles, C. cathayensis were exposed to 0.5 (low exposure, L14) or 1.5 (high exposure, H14) mg/L Cd for 14 days. Our results indicate that Cd exposure leads to oxidative stress and immunosuppression, with the latter effect being larger for exposure to higher Cd concentrations. A total of 2172 differentially expressed genes (DEGs) were identified by transcriptome analysis of the hepatopancreas, of which 885 were upregulated and 1287 were downregulated. Gene ontology and KEGG analyses revealed that the DEGs in the H14 group are enriched for energy generation terms and the "oxidative phosphorylation" pathway, respectively. Therefore, up-regulation of energy metabolism may be an adaptive strategy under Cd stress. Moreover, several genes involved in antioxidant activity were downregulated, whereas genes related to reactive oxygen species generation were upregulated. In addition, many immunity-related genes were identified within the DEGs, indicating that Cd toxicity may affect immune defense. Further, DEGs in the H14 group were enriched for disease-associated pathways. Taken together, our results indicate that Cd exposure leads to metabolic disorders, oxidative stress, and immunosuppression and thus may potentially contribute to disease outbreaks., Competing Interests: Declaration of Competing Interest The authors have declared that no conflict of interest exists., (Copyright © 2023. Published by Elsevier Inc.)

Objective: To explore the sagittal radiological parameters related to clinical recovery of patients with acute traumatic central cord syndrome (ATCCS) and determine the diagnostic value of related variables., Methods: A retrospective review was performed of 104 patients with ATCCS. Six cervical sagittal balance parameters were collected: Cobb angle, T1 slope, neck tilt, thoracic inlet angle (TIA), C2-C7 sagittal vertex axis, T1 slope - C2-C7 Cobb angle. The patients were assigned to an ideal improvement group and poor improvement group according to their recovery rate. Receiver operating characteristic curve and area under the curve were used to evaluate the significant results of logistic regression and the optimal diagnostic value., Results: Preoperative and postoperative Japanese Orthopaedic Association scores indicated a good recovery after surgical intervention. Radiological findings revealed that neck tilt and TIA were risk factors for poor neurological improvement of patients with ATCCS. Area under the curve (95% confidence interval) values of neck tilt and TIA were 0.763 (0.660-0.866) and 0.749 (0.643-0.855), and cutoff values were 39.1° and 65.6°, respectively., Conclusions: Lower neck tilt and TIA are risk factors for poor outcomes in patients with ATCCS after surgery. Neck tilt <39° and TIA <66° had significant diagnostic value for poor prognosis., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Twelve dihydro-β-agarofuran-type sesquiterpenoids, including five new ones (1-5), were purified from the seeds of Celastrus virens (Wang et Tang) C. Y. Chent et T. C. Kao. Their chemical structures were characterized via comprehensive spectroscopic analysis, single-crystal X-ray diffraction analysis, and computational prediction of ECD, as well as comparison of observed and reported NMR spectral data. Among the isolates, nine abundant dihydro-β-agarofuran-type sesquiterpenoids were evaluated for their lifespan-extending activity using the nematode Caenorhabditis elegans model. As a result, compounds 1, 2, 5, 6, 8, and 9 (50 μM) significantly extended the mean survival time of C. elegans, respectively, compared with the blank control group (p < 0.05). Further Quantitative RT-PCR showed that the prolonging of lifespan mediated by compounds 1, 6, 8, and 9 were dependent on the transcription factors skn-1 and hsf-1., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Ovarian cancer is the most lethal gynecologic malignant cancer, frequently due to its late diagnosis and high recurrence. Cancer stem cells (CSCs) from different malignancies including ovarian cancer have been linked to chemotherapy resistance and poor prognosis. Therefore, identifying the molecular mechanisms mediating therapy resistance is urgent to finding novel targets for therapy-resistant tumors. Aberrant O-glycosylation ascribed to subtle alteration of GALNT family members during malignant transformation facilitate metastasis in various cancers. The current study demonstrated that BAG3 was upregulated in platin-resistant ovarian cancer tissues and cells, and high BAG3 predicted dismal disease-free survival of patients with ovarian cancer. In addition, the current study showed that BAG3 facilitated CSC-like properties of ovarian cancer cells via regulation of GALTN10. In a term of mechanism, BAG3 epigenetically regulated GALNT10 transactivation via histone H3 lysine 4 (H3K4) presenter WDR5. We demonstrated that WDR5 increased H3K4 trimethylation (H3K4me3) modification at the promoter regions of GALNT10, facilitating recruitment of transcription factor ZBTB2 to the GALNT10 promoter. Collectively, our study uncovers an epigenetic upregulation of GALNT10 by BAG3 via WDR5 to facilitate CSCs of platin-resistant ovarian cancers, providing additional information for further identification of attractive targets with therapeutic significance in platin-resistant ovarian cancer., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Ovarian cancer is the most frequent cause of gynecologic malignancies associated death. Primary or acquired cisplatin resistance is frequently occurred during ovarian cancer therapy. Cancer stem cells (CSC) tend to form minimal residual disease after chemotherapy and are implicated in relapse. The ability of cancer cells to reprogram their metabolism has recently been related with maintenance of CSC and resistance to chemotherapies. The current study found that BAG5 expression was decreased in cisplatin-resistant ovarian cancer cells and clinical tissues. Our data demonstrated that BAG5 knockdown was implicated in metabolic reprogramming and maintenance of cancer stem cell (CSC)-like features of ovarian cancer cells via regulation of Rictor and subsequent mTORC2 signaling pathway. In addition, the current study demonstrated that Bcl6 upregulation was responsible for repression of BAG5 transactivation via recruitment on the BAG5 promoter in cisplatin-resistant ovarian cancer. The current study also demonstrated reverse correlations between BAG5 and Bcl6, BAG5 and Rictor in ovarian serous adenocarcinoma tissues. Collectively, the current study identified the implication of Bcl6/BAG5/Rictor-mTORC2 signaling pathway in metabolic reprograming and maintenance of CSC-like features in cisplatin-resistant ovarian cancer cells. Therefore, further studies on the mechanism underlying regulation of metabolic reprogramming and CSC-like characteristics of cisplatin-resistant ovarian cancer cells may contribute to the establishment of novel therapeutic strategy for cisplatin-resistance., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Ovarian cancer is the deadliest gynaecologic malignancy, and the five-year survival rate of patients is less than 35% worldwide. Cancer stem cells (CSCs) are a population of cells with stem-like characteristics that are thought to cause chemoresistance and recurrence. TRIM29 is aberrantly expressed in various cancers and associated with cancer development and progression. Previous studies showed that the upregulation of TRIM29 expression in pancreatic cancer is related to stem-like characteristics. However, the role of TRIM29 in ovarian cancer is poorly understood. In this study, we found that TRIM29 expression was increased at the translational level in both the cisplatin-resistant ovarian cancer cells and clinical tissues. Increased TRIM29 expression was associated with a poor prognosis of patients with ovarian cancer. In addition, TRIM29 could enhance the CSC-like characteristics of the cisplatin-resistant ovarian cancer cells. Recruitment of YTHDF1 to m6A-modified TRIM29 was involved in promoting TRIM29 translation in the cisplatin-resistant ovarian cancer cells. Knockdown of YTHDF1 suppressed the CSC-like characteristics of the cisplatin-resistant ovarian cancer cells, which could be rescued by ectopic expression of TRIM29. This study suggests TRIM29 may act as an oncogene to promote the CSC-like features of cisplatin-resistant ovarian cancer in an m6A-YTHDF1-dependent manner. Due to the roles of TRIM29 and YTHDF1 in the promotion of CSC-like features, they may become potential therapeutic targets to combat the recurrence of ovarian cancer., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Cisplatin-based chemotherapies have long been considered as a standard chemotherapy in ovarian cancer. However, cisplatin resistance restricts beneficial therapy for patients with ovarian cancer. The ubiquitin-like protein interferon-stimulated gene 15 (ISG15) encodes a 15-kDa protein, that is implicated in the post-translational modification of diverse proteins. In this work, we found that ISG15 was downregulated in cisplatin resistant tissues and cell lines of ovarian cancer. Functional studies demonstrated that overexpression of wild type (WT) ISG15, but not nonISGylatable (Mut) ISG15 increased cell responses to cisplatin in resistant ovarian cancer cells. Furthermore, we found that WT ISG15 decreased ABCC2 expression at the protein level. Importantly, overexpression of ABCC2 blocked sensitizing effect of ISG15 on cisplatin. In addition, we identified that hnRNPA2B1 was recruited to 5'UTR of ABCC2 mRNA and promoted its translation, which was blocked by ISG15. We further demonstrated that hnRNPA2B1 could be ISGylated, and ISGylation blocked its recruitment to ABCC2 mRNA, thereby suppressed translation of ABCC2. Altogether, our data support targeting ISG15 might be a potential therapeutic strategy for patients with cisplatin-resistant ovarian cancer., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Objective: This study was carried out to analyze the surgical effect of cervical spine sagittal alignment for patients with Hirayama disease (HD)., Methods: Forty-four subjects were retrospectively analyzed for the parameters of cervical spine sagittal alignment. The case group consisted of 23 patients with HD, whereas the control group consisted of 21 healthy adolescent subjects. Pre- and postoperative cervical spine sagittal parameters of the patients with HD were collected; the cervical sagittal parameters of the healthy adolescent subjects were also collected. Sagittal alignment parameters were compared between the patients with HD and the healthy adolescent subjects, and between the pre- and postoperative parameters for the patients with HD., Results: Forty-four subjects completed the follow-up, with the average follow-up period being 18.0 months. No significant differences were detected between the HD and control groups for clinical parameters (P > 0.05). The preoperative HD group had smaller values compared with the control group in the sagittal parameters of C2-7 cervical lordosis (CL) angle, T1 slope, thoracic inlet angle (TIA), and cervical tilt angle (P < 0.05). For the patients with HD, the preoperative values were smaller compared with the postoperative HD values for the parameters of C2-7 CL angle, T1 slope, and cervical tilt angle (P < 0.05). We found no significant differences between the postoperative patients with HD and the healthy subjects, including C2-7 CL angle, C2-7 sagittal vertical axis, T1 slope, TIA, neck tilt angle, cervical tilt angle, and cranial tilt angle (P > 0.05)., Conclusions: Patients with HD have sagittal imbalance of the cervical spine compared with age-matched healthy adolescent subjects, and surgical treatment could correct the sagittal imbalance., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Glucose limitation activates p53, which functions as an adaptive response to maintain cell survival. However, p53 is frequently deleted or mutated in a variety of tumors, while most cancer cells can acclimatize themselves to metabolically unfavorable surrounding, indicating that alternative mechanisms other than p53 transactivation underly adaptive response of cancer cells with p53 deletion or mutation to metabolically hostile environment. Sestrin 2 (SESN2) is a p53 downstream target, which plays a protective role against various stressful stimuli, such as genotoxic, energetic, and oxidative stress. In the current study, we demonstrated that SESN2 transcript was stabilized by glucose limitation at the posttranscriptional level irrespective of p53 status. Importantly, SESN2 also protected cells from metabolic stress triggered by glucose limitation in a p53-independent manner. Our data indicated that stabilization of SESN2 transcript might be an alternative adaptive response to metabolic stress other than p53 activation. Thereby, the current study highlights the significance of stabilization of SESN2 transcript in adaptation of cells with p53 deletion or mutation to metabolic stress., (Copyright © 2019 Elsevier Inc. All rights reserved.)

BAG3 is a member of the cochaperone BAG family and often highly expressed in various cancers. Recently, evidences show that BAG3 promotes stemness of human cancer cells. IFN-stimulated genes 15 (ISG15) is an ubiquitin-like molecule, which is covalently conjugated with substrates to form ISGylated proteins. Global screening BAG3 interacting partners demonstrated that ISG15 might be a potential binding partner. The current study revealed that BAG3 did not interact with ISG15, but positively regulated ISG15 expression in pancreatic ductal adenocarcinoma cancer (PDAC). It was further found that BAG3 deletion stabilized ISG15 mRNAs, while suppressed its translation via increasing Serine phosphorylation of Ago2 at position 387 (S387). Both BAG3 deletion and ISG15 knockdown suppressed stem cell-like phenotypes of PDAC cells, including clonogenicity, invasiveness and spheroid formation. In addition, ectopic ISG15 expression rescued the suppressive role of BAG3 deletion in cancer stem cell (CSC)-like phenotypes of PDAC cells, and this effect of ISG15 was independent of its ISGylation function. The current study implies that BAG3 and ISG15 may provide a therapeutic advantage for PDAC., (Copyright © 2019. Published by Elsevier B.V.)

Background: Spinal cord injury may cause cortical reconstruction. We, therefore explored the changes in cortical activation before and after anterior cervical decompression and fusion surgery in patients with Hirayama disease (HD)., Methods: In total, 17 cases with HD underwent anterior cervical decompression and fusion surgery. Blood oxygenation level-dependent functional magnetic resonance imaging scan was performed preoperatively, 3 months, 6 months, and 1 year after surgery. Activated voxels were compared between both hands after adjusting for head motion, slice timing, spatial normalization, and image smoothing. Grip strength also was tested in both hands., Results: A retrospective review indicated that the grip strength of the asymptomatic hand was significantly stronger than the symptomatic hand at the time point before the surgery, 3 months after surgery, 6 months after surgery, and 1 year after surgery (P < 0.001). The grip strength of both symptomatic and asymptomatic hands continuously increased within 6 months after surgery (P < 0.05), but it stopped at 1 year after the surgery. The symptomatic limb tends to produce bilateral activation in the primary motor area (M1) during motor tasks. Both contralateral and ipsilateral M1 activation were stronger in symptomatic hand tasks preoperatively (P < 0.05). Both contralateral and ipsilateral activation in M1 during symptomatic hand tasks began to reduce after surgery, and statistical significance was observed 6 months after surgery (P < 0.05). Contralateral activation was relatively even over 6 months of the surgery (P > 0.05)., Conclusions: After surgery, pathologic reconstruction may have occurred in the primary motor cortex. Recovery of motor function in the symptomatic limb was accompanied by decreased ipsilateral and contralateral M1 activation, as well as symptom improvement. These findings suggested that postoperative cortical activation changes may reflect functional recovery in HD., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Background: To explore the predictive parameters for adjacent segment disease (ASD) after anterior cervical arthrodesis at the sagittal measurement of thoracic inlet., Methods: We included 212 subjects treated with anterior cervical fusion surgery to identify predictive parameters for ASD. We applied multivariate logistic regression to find the relevant parameters., Results: Two hundred patients completed the final follow-up. The results of logistic regression analysis showed that T1 slope of less than 19.50 could be a predictive parameter of ASD after the anterior cervical arthrodesis (P < 0.05)., Conclusions: Sagittal balance of thoracic inlet is vital to avoid the. cervical degenerative disease, while T1 slope of less than 19.50 appears to be an independent risk factor for the ASD, keeping the T1 slope of more than 19.5 is reasonable., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Stabilization/Solidification (S/S) can be regarded as necessary for remediation of heavy metal contaminated soil. There is, however, solid agent is not very convenient to use. Water-soluble thiourea-formaldehyde (WTF) is a novel chelating agent, which has more practical applications. The process of WTF resin for S/S process of heavy metal contaminated soils was studied. Laboratory-prepared slurries, made of field soils spiked with Cd 2+ and Cr 6+ were treated with WTF resin. The toxicity characteristic leaching procedure (TCLP) showed that with 2 wt% WTF, in the neutral condition of soil after treatment for 7 d, the leaching concentrations of Cd 2+ and Cr 6+ in contaminated soil were decreased by 80.3% and 92.6% respectively. Moreover, Tessier sequence extraction procedure showed WTF resin reduced the leaching concentration by transforming heavy metal from exchange form to organic form. The structure of WTF is obtained according to elemental analysis result and reaction mechanism. Through analysis of the infrared spectrogram of WTF and WTF heavy mental chelating precipitation, WTF can form stable chelate with heavy mental through coordination. The significant groups are hydroxyl, nitrogen and sulphur function groups in WTF mainly. Toxicology test revealed that the WTF resin is nontoxic to microorganism in the soils., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Posttranscriptional regulation process plays important roles in renal disease pathogenesis. AU-rich element RNA-binding protein (AUF1) interacts with and destabilizes mRNAs containing AU-rich elements (AREs) in their 3'UTR. The current study demonstrated that AUF1 was increased in unilateral ureteral obstruction (UUO) animal models. While proliferation and migration of HK2 cells was unaltered by AUF1 downregulation under normal condition, proliferative inhibition and migratory promotion mediated by TGF-β was significantly compromised. Mechanically, AUF1 downregulation decreased phosphorylated Smad2/3 via increasing their E3 ligase Nedd4L at the posttranscriptional level. In addition, the current study identified Nedd4L as a previously unreported target of AUF1. AUF1 regulates Nedd4L expression at the posttranscriptional level by interaction with AREs in the 3'UTR of the Nedd4L mRNA. Collectively, the current study indicates that AUF1 might be a potential player in renal tubulointerstitial fibrosis through modulation of TGF-β signal transduction via posttranscriptional regulation of Nedd4L., (Copyright © 2017. Published by Elsevier B.V.)

Bcl-2 associated athanogene 3 (BAG3) contains a modular structure, through which BAG3 interacts with a wide range of proteins, thereby affording its capacity to regulate multifaceted biological processes. BAG3 is often highly expressed and functions as a pro-survival factor in many cancers. However, the oncogenic potential of BAG3 remains not fully understood. The cell cycle regulator, S-phase kinase associated protein 2 (Skp2) is increased in various cancers and plays an important role in tumorigenesis. The current study demonstrated that BAG3 promoted proliferation of ovarian cancer cells via upregulation of Skp2. BAG3 stabilized Skp2 mRNA via its 3'-untranslated region (UTR). The current study demonstrated that BAG3 interacted with Skp2 mRNA. In addition, miR-21-5p suppressed Skp2 expression, which was compromised by forced BAG3 expression. These results indicated that at least some oncogenic functions of BAG3 were mediated through posttranscriptional regulation of Skp2 via antagonizing suppressive action of miR-21-5p in ovarian cancer cells., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Objective: To explore risk factors affecting surgical results of Hirayama disease., Methods: A retrospective analysis of 210 patients was performed to identify risk factors affecting surgical results of Hirayama disease by using univariate and multivariate analyses. A receiver operating characteristic curve and area under the curve were applied to evaluate the significant results of the multivariate analysis and the optimal reference value., Results: The mean follow-up period was 27.3 months (range, 14-45 months), and 194 patients with clinical and radiographic data completed the final follow-up. Multivariate analysis identified age of patients (cutoff value 22.5 years), duration of the disease (cutoff value 33 months), physiologic reflex, and pathologic reflex as independent risk factors for surgical results of Hirayama disease. The receiver operating characteristic curve analysis and area under the curve showed that good reference value was obtained for the risk factors., Conclusions: Age of patient, duration of the disease, physiologic reflex, and pathologic reflex are the main risk factors affecting surgical results of Hirayama disease. Receiver operating characteristic analysis shows that good reference value was obtained for the risk factors., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Optical reporters for cAMP represent a fundamental advancement in our ability to investigate the dynamics of cAMP signaling. These fluorescent sensors can measure changes in cAMP in single cells or in microdomains within cells as opposed to whole populations of cells required for other methods of measuring cAMP. The first optical cAMP reporters were FRET-based sensors utilizing dissociation of purified regulatory and catalytic subunits of PKA, introduced by Roger Tsien in the early 1990s. The utility of these sensors was vastly improved by creating genetically encoded versions that could be introduced into cells with transfection, the first of which was published in the year 2000. Subsequently, improved sensors have been developed using different cAMP binding platforms, optimized fluorescent proteins, and targeting motifs that localize to specific microdomains. The most common sensors in use today are FRET-based sensors designed around an Epac backbone. These rely on the significant conformational changes in Epac when it binds cAMP, altering the signal between FRET pairs flanking Epac. Several other strategies for optically interrogating cAMP have been developed, including fluorescent translocation reporters, dimerization-dependent FP based biosensors, BRET (bioluminescence resonance energy transfer)-based sensors, non-FRET single wavelength reporters, and sensors based on bacterial cAMP-binding domains. Other newly described mammalian cAMP-binding proteins such as Popdc and CRIS may someday be exploited in sensor design. With the proliferation of engineered fluorescent proteins and the abundance of cAMP binding targets in nature, the field of optical reporters for cAMP should continue to see rapid refinement in the coming years., (Published by Elsevier Ltd.)

Objective: The role of autophagy in moderate hypothermia in posttraumatic brain injury (post-TBI) remains elusive. In this study, we evaluated the protective role of autophagy in post-TBI moderate hypothermia., Methods: Adult male Sprague-Dawley rats were randomly divided into 3 groups (n = 36/group): TBI with hypothermia group (sham), TBI with hypothermia and a single intracerebroventricular injection of saline (saline, 5 μL), and TBI with hypothermia and a single intracerebroventricular injection of 3-methyladenine (600 nmol, diluted in 0.9% saline to a final volume of 5 μL). All rats, except those in the behavioral tests, were killed at 24 hours after fluid percussion TBI. Immunohistochemistry staining, western blot, and transmission electron microscopy were performed to assess changes in apoptosis and autophagy after injection of 3-methyladenine. Motor function (beam-walk test) and spatial learning/memory (Morris water maze) were assessed on postoperative days 1-5 and 11-15, respectively., Results: Our results showed downregulation of the expression level of microtubule-associated protein 1 light chain 3 and Beclin-1, aggravation of behavioral outcome, and increase of apoptosis., Conclusion: Our results suggest that the autophagy pathway is involved in the neuroprotective effect of post-TBI hypothermia and negative modulation of apoptosis may be 1 possible mechanism., (Copyright © 2016. Published by Elsevier Inc.)

Objective: Multiple mechanisms participated in the cell death after fluid percussion traumatic brain injury (TBI). In the present study, we evaluated the effect on cell death in the injured cortex after fluid percussion TBI and investigated a possible role of autophagy., Methods: TBI model was induced by a fluid percussion TBI device. Moderate hypothermia (32°C) was achieved by partial immersion in a water bath (0°C) under general anesthesia for 4 hours. All rats were killed at 6 or 24 hours after TBI., Results: Cleaved caspase 3 evaluated with Western blotting and terminal deoxynucleotidyl transferase-mediated 2'-deoxyuridine 5'-triphosphate-biotin nick end labeling-positive cells in injured cortex were significantly increased 6 hours after fluid percussion TBI and were further up-regulated 24 hours after TBI, dramatic up-regulation of Beclin-1 and protein light chain-3 expression levels was also observed. Further up-regulation of biomarkers of autophagy, attenuation of caspase 3 up-regulation and reduction of cell death was observed after 4 hours of hypothermia. Immunofluorescence analysis for cell localization demonstrated that protein light chain-3- and Beclin-1-positive cells included neurons and glial cells in the injured cortex after TBI and hypothermic treatment. By ultrastructural observation, autolysosomes in injured cortex were significantly increased at 6 and 24 hours after TBI and were further up-regulated after TBI hypothermic treatment., Conclusions: These data suggest that hypothermic treatment could attenuate TBI-induced cell death in this fluid percussion TBI model, possibly through activation of autophagy pathway., (Copyright © 2015 Elsevier Inc. All rights reserved.)

The current global strategy for the control of soil-transmitted helminthiasis emphasises periodic administration of anthelminthic drugs to at-risk populations. However, this approach fails to address the root social and ecological causes of soil-transmitted helminthiasis. For sustainable control, it has been suggested that improvements in water, sanitation and hygiene behaviour are required. We designed a 5-year multi-intervention trial in Menghai county, Yunnan province, People's Republic of China. Three different interventions were implemented, each covering a village inhabited by 200-350 people. The interventions consisted of (i) initial health education at study inception and systematic treatment of all individuals aged ≥2 years once every year with a single dose of albendazole; (ii) initial health education and bi-annual albendazole administration; and (iii) bi-annual treatment coupled with latrine construction at family level and regular health education. Interventions were rigorously implemented for 3 years, whilst the follow-up, which included annual albendazole distribution, lasted for 2 more years. Before the third round of treatment, the prevalence of Ascaris lumbricoides was reduced by only 2.8% in the annual treatment arm, whilst bi-annual deworming combined with latrine construction and health education resulted in a prevalence reduction of 53.3% (p<0.001). All three control approaches significantly reduced the prevalence of Trichuris trichiura and hookworm, with the highest reductions achieved when chemotherapy was combined with sanitation and health education. The prevalence of T. trichiura remained at 30% and above regardless of the intervention. Only bi-annual treatment combined with latrine construction and health education significantly impacted on the prevalence of Taenia spp., but none of the interventions significantly reduced the prevalence of Strongyloides stercoralis. Our findings support the notion that in high-endemicity areas, sustainable control of soil-transmitted helminth infections necessitates measures to reduce faecal environmental contamination to complement mass drug administration. However, elimination of soil-transmitted helminthiasis will not be achieved in the short run even with a package of interventions, and probably requires improvements in living conditions, changes in hygiene behaviour and more efficacious anthelminthic drugs and treatment regimens., (Copyright © 2014 Elsevier B.V. All rights reserved.)

The presence of large Trichuris spp. eggs in human faecal samples is occasionally reported. Such eggs have been described as variant Trichuris trichiura or Trichuris vulpis eggs. Within the frame of a randomised controlled trial, faecal samples collected from 115 Bulang individuals from Yunnan, People's Republic of China were subjected to the Kato-Katz technique (fresh stool samples) and the FLOTAC and ether-concentration techniques (sodium acetate-acetic acid-formalin (SAF)-fixed stool samples). Large Trichuris spp. eggs were noted in faecal samples with a prevalence of 6.1% before and 21.7% after anthelminthic drug administration. The observed prevalence of standard-sized T. trichiura eggs was reduced from 93.0% to 87.0% after treatment. Considerably more cases of large Trichuris spp. eggs and slightly more cases with normal-sized T. trichiura eggs were identified by FLOTAC compared to the ether-concentration technique. No large Trichuris spp. eggs were observed on the Kato-Katz thick smears., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Objective: The Chinese Head Trauma Data Bank (CHTDB) has been established, which includes 7,145 hospitalised cases with acute head trauma patients in 47 hospitals., Methods: We explored factors that might affect the outcome of acute traumatic brain injury., Results: There was no statistical difference in the mortality rate between male (7.5%) and female (7.2%) patients (P>0.05). The mortality rate in children (<18 years), adults (18-65 years) and elderly (>65 years) was 7.3%, 7.2% and 9.0%, respectively (P>0.05). The mortality rate of patients with mild (2.7%), moderate (5.0%) and severe (21.8%) head trauma was significantly different (P<0.001). The mortality rate of patients with unilateral tentorial herniation, bilateral tentorial herniation and tonsillar herniation was 24.2%, 60.2% and 78.8% respectively (P<0.001). The mortality rate of patients with intracranial pressure (ICP)<20 mm Hg, 20-40 mm Hg and >40 mm Hg was 6.3%, 21.4% and 93.1%, respectively (P<0.001). The mortality rate of patients with no cerebral contusion, single cerebral contusion and multiple cerebral contusions was 3.9%, 7.8% and 14.8%, respectively (P<0.001). The mortality rate of patients with and without traumatic subarachnoid haemorrhage (tSAH) was 9.5% and 5.4%, respectively (P<0.001). The mortality rate of patients with no intracranial haematomas, single intracranial haematoma and multiple intracranial haematomas was 5.8%, 8.4% and 20.6%, respectively (P<0.001)., Conclusion: The CHTDB, the first head trauma data bank in China, has one of the largest numbers of cases of any head trauma data bank in the world. Our major findings on mortality may be helpful to neurosurgeons for predicting the outcome of acute head trauma patients., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Monocyte chemoattractant protein-1 (MCP-1) plays crucial roles at multiple stages of atherosclerosis. We hypothesized that MCP-1 might impair the reverse cholesterol transport (RCT) capacity of HepG2 cells by decreasing the cell-surface protein expression of ATP binding cassette A1 (ABCA1), ATP binding cassette G1 (ABCG1), and scavenger receptor class B type I (SR-BI). MCP-1 reduced the total protein and mRNA levels of ABCA1 and SR-BI, but not of ABCG1. MCP-1 decreased the cell-surface protein expression of ABCA1, ABCG1, and SR-BI in dose-dependent and time-dependent manners, as measured using cell-surface biotinylation. We further studied the phosphoinositide 3-kinase (PI3K)/serine/threonine protein kinase Akt pathway in regulating receptor trafficking. Both the translation and transcription of ABCA1, ABCG1, and SR-BI were not found to be regulated by the PI3K/Akt pathway. However, the cell-surface protein expression of ABCA1, ABCG1, and SR-BI could be regulated by PI3K activity, and PI3K activation corrected the MCP-1-induced decreases in the cell-surface protein expression of ABCA1, ABCG1, and SR-BI. Moreover, we found that MCP-1 decreased the lipid uptake by HepG2 cells and the ABCA1-mediated cholesterol efflux to apoA-I, which could be reversed by PI3K activation. Our data suggest that MCP-1 impairs RCT activity in HepG2 cells by a PI3K/Akt-mediated posttranslational regulation of ABCA1, ABCG1, and SR-BI cell-surface expression.

Dissolved organic matter (DOM) is attributed a key role in soil major biogeochemical processes. Its molecular characteristics can reflect both its source and its biogeochemical history. Saline-alkali soil samples were collected from a field in desert and semi-desert region. DOM was analyzed by synchronous fluorescence spectroscopy. Principal Components Analysis (PCA) of synchronous fluorescence spectra was used to detect and describe changes in the DOM in different soil profiles. The analysis of PCA yielded 2 PCs that account for 97% of the variance. DOM is characterized by a dominant tyrosine-like fluorophore in the Region I (PC1>0.9, PC2<0.3), including soils Ke02, Ke03, Ke04, Rh02, Rh03, Rh04, Sa04 and Sc04. The region II including Kf02, Sa01, Yb01, Yzd01 and Yzd02 obtain higher PC2 (>0.9) and lower PC1 (<0.3) loading, which is dominated humic-like fluorophore. Hierarchical cluster analysis (HCA) identified five main clusters. The results of hierarchical cluster analysis (HCA) were consistent with the PCA. The ratio of fluorescence intensity of humic substances at long wavelengths and short wavelengths was used to indicate the humification of organic matter. Two ratios (I(345/318) and I(362/318).) can be used to indicate the humification of DOM. The humification degree of the remaining soils has no systematic trend because of land use, agricultural irrigation, soil erosion and salinization. Soils obtained a relatively high humification degree in the region II and a relatively low humification degree in the region I., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Dissolved organic matter (DOM) was extracted from six sediment samples in arid and semi-arid region, which was characterized by fluorescence excitation-emission matrices (EEMs). The results showed that four fluorescent peak, fulvic-like (peak A), humic-like (peak C) and two tryptophan-like (peaks B and D), were identified in lake sediment DOM. Fluorescence quenching titration showed that peaks B and D were quenched gradually by adding additional Cu (II) and Hg (II), whereas humic-like substances had no systematic trend of the change of fluorescence intensity. Increasing fluorescence intensity value of humic-like substances can also be found. The modified Stern-Volmer model was used to calculate conditional stability constants (logK) and the percent of fluorophores (f %) which participate in the complexation between DOM and Cu (II), and Hg (II). The results showed that DOM-Cu (II) and DOM-Hg (II) complexes had higher logK values of 4.21-5.23 and the logK values of DOM-Cu (II) are much larger than the corresponding values for Hg (II). Peak B showed relatively low logK and high f % values than those of peak D. Different pollution sources which are mainly obtained from the upstream industrial wastewater, domestic sewage and return water of farmland irrigation tend to affect the stability constants and complexing capacities of Cu (II) and Hg (II)., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Objective: Rebleeding from ruptured intracranial aneurysms is a major cause of death and disability. The aim of this study was to investigate the incidence of rebleeding and the risk factors related to rebleeding before early aneurysm repair., Methods: The incidence of rebleeding, demographic data, and clinical data from 326 patients with aneurysmal subarachnoid hemorrhage (SAH) were retrospectively collected. All clinical variables were examined by univariate analysis, and a binary logistic regression analysis was performed to identify the risk factors related to rebleeding., Results: Rebleeding occurred in 70 (21.5%) of the 326 aneurysm patients during transfer or during the in-hospital stay (within 72 hours); 24 episodes (34.3%) occurred within 3 hours, and 44 episodes (62.9%) occurred within 6 hours after the initial SAH. Univariate analysis showed that there were significant differences between the rebleeding and nonrebleeding patients in terms of age, aneurysm size, systolic arterial blood pressure (SBP), Hunt-Hess grade and outcome at discharge. The binary logistic regression analysis revealed that age (odds ratio [OR] = 1.167), aneurysm size (OR = 1.624), SBP (OR = 3.338), and Hunt-Hess grade (OR = 2.512) were independent risk factors for aneurysmal rebleeding (for each P < 0.05)., Conclusions: The incidence of early aneurysmal rebleeding within hours after the initial SAH is high during transfer or during the in-hospital stay. Advanced age, an aneurysm size larger than 10 mm, SBP higher than 160 mmHg, and poor Hunt-Hess grade were independent risk factors for aneurysmal rebleeding. The importance of early aneurysm repair should be emphasized because aneurysmal rebleeding contributes to a poor outcome., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Objective: The H-reflex on stimulation of the tibial nerve in the popliteal fossa is routinely used in the diagnosis of first sacral (S1) nerve-root radiculopathy. The H-reflex latency, however, is considered to lack sensitivity since a small change from the focal root pathology can be diluted in a relatively long reflex latency. We have studied the soleus H-reflex elicited by stimulation of the S1 nerve root at the S1 foramen. The normal values for the S1-foramen H-reflex have been reported in a previous study, but there are no definitive reports in patients with S1 radiculopathy. This study was undertaken to determine whether stimulating at the S1 nerve root can improve the utility of the H-reflex for detecting an S1-root lesion., Methods: A randomised paired-study design was utilised to evaluate two H-reflexes: one elicited with tibial nerve stimulation and one elicited with S1-root stimulation. Fifty-five patients with unilateral S1 radiculopathy, confirmed by clinical, electrodiagnostic and magnetic resonance imaging (MRI) evidences were studied. A high-voltage electrical stimulator was used to elicit H-reflexes bilaterally at the S1 foramen and L4/L5 spine level. Latencies were compared with previously generated normal values and similar responses from the asymptomatic leg, focussing on the interval between the peak of M- and H-waves (HMI)., Results: On the symptomatic side, 39 of the 55 patients had abnormal tibial H-reflex latencies and 54 patients had abnormal responses on S1-foramen stimulation (absent in 18; HMI prolonged >0.4ms in 36). On the asymptomatic side, all 55 patients had normal tibial H-reflexes, and 52 had normal responses on S1-foramen stimulation. In three patients, the HMI was abnormal on S1-foramen stimulation. In 46 patients tested with L4/L5-level stimulation, H-reflex was present in 39 and absent in seven. The latency of the M-wave to S1 stimulation was normal., Conclusions: Abnormal S1-root H-reflexes reveal lesions at the S1 root in patients with normal tibial H-reflexes; therefore, enhancing diagnostic sensitivity. The appearance of the H-reflex to L4/L5-level stimulation in patient with absent H-reflex to S1-foramen stimulation further localises the site of S1 nerve-root lesion to the L5/S1 spine level. Thus, H-reflex to S1-root stimulation significantly increases the diagnostic sensitivity for S1 radiculopathy., Significance: In our study, the S1-root H-reflex with high-voltage electrical stimulation has shown greater sensitivity than the tibial H-reflex in evaluating S1 compressive radiculopathies. An abnormal S1-root H-reflex helps to localise the lesion to the S1 root in patients with concurrent abnormal tibial nerve H-reflex, which may increase diagnostic specificity., (Copyright 2010 International Federation of Clinical Neurophysiology. Published by Elsevier Ireland Ltd. All rights reserved.)

Background: To explore the dose-response effects of topical administration of nimodipine on cerebral vasospasm (CVS) after subarachnoid hemorrhage (SAH) in rabbits., Methods: The CVS model was established by injection of fresh autologous nonheparinized arterial blood into the subtemporal area of basilar cisterns. The 24 CVS animals were randomly divided into 4 groups, group I (n=7): nimodipine original stock solution/normal saline=1/19 (0.01 mg/mL); group II (n=6): nimodipine original stock solution/normal saline=1/9 (0.02 mg/mL); group III (n=5): nimodipine original stock solution/normal saline=1/4 (0.04 mg/mL); and group IV (n=6) with no nimodipine, but 5% ethanol dissolved in normal saline as the control group. The operative area was administrated with nimodipine at different concentrations or alcohol-saline at 3 days after SAH. The blood flow velocity of middle cerebral artery was measured at 5, 15, 30, and 60 minutes after topical administration of nimodipine by transverse cerebellar diameter monitoring., Results: Blood flow velocity of middle cerebral artery in group II (0.02 mg/mL) and in group III (0.04 mg/mL) significantly decreased at 60 and 15 minutes, respectively, after topical administration of nimodipine (P<0.05), and even more significantly at 30 and 60 minutes after topical administration of nimodipine in group III (0.04 mg/mL) (P<0.01)., Conclusion: Topical administration of nimodipine at the concentrations of 1:5 (0.04 mg/mL) and 1:10 (0.02 mg/mL) significantly alleviates CVS after SAH, which indicates that topical administration of nimodipine may be useful for CVS of patients with SAH during surgical clip of intracranial aneurysms.

The purpose of this study was to improve our understanding of the molecular epidemiology of human Blastocystis, focusing on 239 randomly selected individuals in a single village in Yunnan province, China. Emphasis was placed on the relative frequency of different Blastocystis subtypes and underlying risk factors. We used a cross-sectional study design, by employing a pre-tested questionnaire to obtain demographic data and behavioural risk factors, and collected faecal samples for culture and subsequent identification of Blastocystis. DNA was extracted from Blastocystis isolates and the subtypes were identified using 7 subtype-specific sequenced-tagged site (STS) primers. Overall, 78 faecal samples were Blastocystis culture-positive (32.6%, 95% confidence interval: 26.7-38.6%). The majority (n=73, 93.6%) were single infections with one of the known subtypes, whereas 2 isolates consisted of 2 concurrent subtypes. The remaining 3 isolates could not be identified with the currently known STS primers. Risk factors for a Blastocystis infection were drinking unboiled water, consumption of raw water plants and pig ownership. The consumption of raw water plants was positively associated with subtype 1 infections, and drinking unboiled water with subtype 3 infections. In conclusion, human Blastocystis was common in this village in southwest China, and different subtypes were associated with distinct transmission routes or sources of infection, and hence Blastocystis subtypes might be linked to specific environmental compartments.

It has been shown that the stromal-cell population found in bone marrow can be expanded and differentiated into cells with the phenotypes of bone, cartilage, muscle, neural, and fat cells. However, whether mesenchymal stem cells (MSCs) are present in human umbilical-cord blood (UCB) has been the subject of ongoing debate. In this study, we report on a population of fibroblastlike cells derived from the mononuclear fraction of human UCB with osteogenic and adipogenic potential, as well as the presence of a subset of cells that have been maintained in continuous culture for more than 6 months. These cells were found to express CD29, CD44, CD90, CD95, CD105, CD166, and MHC class, but not CD14, CD34, CD40, CD45, CD80, CD86, CD117, CD152, or MHC class II. We also compared gene expression after gene transfer using lenti- and adenoviral vectors carrying the green fluorescence protein to the MSCs derived from UCB because a reliable gene-delivery system is required to transfer target genes into MSCs, which have attracted attention as potential platforms for the systemic delivery of therapeutic genes. The lentiviral vectors can transduce these cells more efficiently than can adenoviral vectors, and we maintained transgene expression for at least 5 weeks. This is the first report showing that UCB-derived MSCs can express exogenous genes by way of a lentivirus vector. These results demonstrate that human UCB is a source of mesenchymal progenitors and may be used in cell transplantation and a wide range of gene-therapy treatments.

Hyperlipidemia, a condition normally observed in cholestatic liver disease, is also a risk factor for the development of atherosclerosis. The relationship between the elevation of lipoproteins in cholestatic liver diseases and atherosclerosis formation has not been elucidated. In this study, we propose that the impairment of endothelium-dependent relaxation (EDR) of blood vessels in cholestatic liver diseases may lead to the development of atherosclerosis. Using bile duct ligation (BDL) in rats as a model, we examined the liver function, serum lipid profile, EDR and morphologic change of the aorta from both sham operated and BDL rats. Significant increases in liver and spleen weights, serum alanine transaminase (ALT) and aspartate transaminase (AST) activities and the bilirubin level were observed in BDL rats. Upon bile duct ligation, the total and low-density lipoprotein cholesterol levels were increased but the high-density lipoprotein cholesterol and triglyceride levels were reduced. Less contractility and lowered response to acetylcholine-induced relaxation were found in aorta segments. In addition, the acetylcholine-induced relaxation was blocked by both L-NAME and 15 mM KCl. Our results suggest that both nitric oxide and endothelium-derived hyperpolarizing factor are important elements for the impairment of the EDR in BDL rats. In addition, a mild atrophy of the media of the aorta was detected in BDL rats. We conclude that the alterations of lipid profile and the mild atrophy of the media may lead to the impairment of EDR in the aorta in BDL rats, and these factors may potentiate the development of atherosclerosis.

Vitrification of oocytes and embryos has recently been improved using new physical supports such as electron microscope (EM) grids, open-pulled straws, and cryoloops. However, the number of samples per container was restricted in each of these methods. In the present study, to develop a novel simple technique for vitrification of large quantities of oocytes or embryos, we examined vitrification of large quantities of immature bovine oocytes using nylon mesh as a novel container. As many as 65 oocyte-cumulus cell complexes could be placed on nylon mesh for vitrification compared with 15 per EM grid. Recovery rates were higher when using nylon mesh than EM grids, while fertilization and development rates were not different. These results indicated that vitrification using nylon mesh is useful and offers a new way to cryopreserve large numbers of oocytes.

Lysophosphatidylcholine (lyso-PC) and arachidonate are products of phosphatidylcholine hydrolysis by phospholipase A(2). In this study, the modulation of arachidonate release by exogenous lyso-PC in rat heart myoblastic H9c2 cells was examined. Incubation of H9c2 cells with lyso-PC resulted in an enhanced release of arachidonate in both a time- and dose-dependent fashion. Lyso-PC species containing palmitoyl (C(16:0)) or stearoyl (C(18:0)) groups evoked the highest amount of arachidonate release, while other lysophospholipid species were relatively ineffective. Cells treated with phospholipase A(2) inhibitors resulted in the attenuation of the enhanced arachidonate release in the presence of lyso-PC. Lyso-PC caused the translocation of phospholipase A(2) from the cytosol to the membrane fraction and induced an increase in Ca2+ flux from the medium into the cells. Nimodipine, a specific Ca(2+)-channel blocker, partially attenuated the lyso-PC-induced rise in intracellular Ca2+. Concurrent with Ca2+ influx, lyso-PC caused an enhancement of protein kinase C activity. The lyso-PC-induced arachidonate release was attenuated when cells were pre-incubated with specific protein kinase C and mitogen activated protein kinase kinase inhibitors. Taken together, these results strongly indicate that the lyso-PC-induced increases in levels of intracellular calcium and stimulation of protein kinase C lead to the activation of cytosolic phospholipase A(2) which results in the enhancement of arachidonate release in H9c2 cells.

Two-cell embryos derived from immature rdw rats by in vitro fertilization (IVF) were vitrified in ethylene glycol-based solutions. Embryos exposed to EFS20 before being vitrified in EFS40 exhibited improved viability in vitro. All embryos exposed to EFS20 for 1-3 min before vitrification in EFS40 were morphologically normal. However, 2-3 min of exposure to EFS20 increased the number of embryos that developed beyond the four-cell stage. More embryos exposed to EFS20 for 2-3 min developed to morulae (63-64%) and blastocysts (34-38%) than those exposed for 1 min (35 and 10%, respectively). After transfer, 33% of embryos exposed to EFS20 for 3 min and vitrified in EFS40 developed to term compared to 29% of fresh embryos. Fifteen (47%) of live young were homozygous rdw and all of the others were heterozygous rats. The present study demonstrated that vitrification in EFS solution can be routinely used to cryopreserve rat two-cell IVF-embryos with no loss of viability., (Copyright 1999 Academic Press.)

Background: Hotien county in Xinjiang province, China, is an area of severe iodine deficiency and has a high infantmortality rate. We investigated whether iodine replacement through iodination of the irrigation water would decrease infant mortality., Methods: We added potassium iodate to irrigation water over a 2 to 4 week period beginning in 1992 in certain areas of three townships (Tusala, Long Ru, and Bakechi). Logistic regression analysis was used to compare the odds ratios for infant and neonatal mortality in treated and intreated areas., Findings: The median urinary iodine concentration significantly increased in women of child-bearing age from < 10 micrograms/L to 55 micrograms/L. Infant-mortality rates decreased in the treated areas of Long Ru (mean [SD] 58.2 [4.4] per 1000 births to 28.7 [7.1] per 1000 births), Tusala (47.4 [12.4] per 1000 births to 19.1 [1.5] per 1000 births), and Bakechi (106.2 [9.5] per 1000 births to 57.3 [7.3] per 1000 births). Similar results were also seen for neonatal mortality. On regression analysis iodine treatment and time were significant independent predictors of infant mortality., Interpretation: Iodine supplementation of irrigation water in areas of severe iodine deficiency decreases neonatal and infant mortality. Iodine replacement has probably been an important factor in the national decrease in infant mortality in China.

We examined the effects of an exogenous mu opioid agonist and antagonist on systemic physiology and neurological outcome following TBI in the rat. Experiment I: [D-Ala2,N-Me-Phe4,Gly5-ol]-enkephalin (DAMGO) (0.1 nMol or 0.3 nMol in 5 microliters) (n = 10) or artificial CSF (n = 10) was administered 5 min prior to fluid-percussion brain injury (2.1 atmospheres). Motor performance was assessed on days 1-5 after TBI. The mu receptor agonist, DAMGO significantly reduced both beam-walking latency and body weight loss after injury (p < 0.05). DAMGO-treated rats (n = 5) did not differ from CSF-treated rats (n = 5) on either systemic arterial blood pressure or heart rate responses to injury. Experiment II: Beta-funaltrexamine (beta-FNA) (20.0 nMol in 5.0 microliters) (n = 10) or artificial CSF (n = 10) was administered (icv) to rats 5 min prior to fluid-percussion brain injury (1.8 atmospheres). Motor performance was assessed on days 1-5 after TBI. The mu receptor antagonist, beta-FNA, significantly increased beam-walking latency after injury (p < 0.05). beta-FNA-treated rats (n = 5) did not differ from CSF-treated rats (n = 5) on either systemic arterial blood pressure or heart rate responses to injury. Experiment III: Neither beta-FNA nor DAMGO affected motor performance in uninjured rats. These results suggest that activation of mu opioid receptors by exogenous agonists may provide protection against deficits in motor performance produced by fluid percussion brain injury.