[Bioinformatics database and production layout visual analysis platform of medicinal plants:a case study of Dao-di medicinal materials in Sichuan province].

In the new stage of trans-omics and trans-subjects for medicinal plants, it is an urgent need to integrate big data, provide interactive applications, and form a unified and multi-level research system and big data platform. Dao-di medicinal material, as an important source of medicinal plants, is a unique quality concept and comprehensive standard of tranditional Chinese medicine(TCM). Several databases have been developed in China and abroad, such as the Encyclopedia of Traditional Chinese Medicine(ETCM) and the Global Pharmacopoeia Genome Database(GPGD). Yet, most databases do not provide multi-dimensional data, including geographic data, phenotype data, compound data, and genetic data. Sichuan, known as the hometown of TCM therapies and the treasure trove of TCM, is the most representative region of medicinal plant diversity in China. According to the latest data of the fourth national survey of TCM resources, there are more than 8 000 TCM and 86 Dao-di medicinal materials in Sichuan province. Based on resource census data and relevant achievements, this study constructed the bioinformatics database of medicinal plants and the visual analysis platform of production layout by taking the Dao-di medicinal materials in Sichuan province as an example, covering geographic data, phenotype data, compound data, and genetic data. It effectively integrates multi-dimensional data of Dao-di medicinal materials and provides different levels of data interaction applications. The platform is the first large-scale multi-dimensional database and visual platform of Dao-di medicinal materials in Sichuan province, which serves as an essential resource for germplasm resources identification, decomposition of biosynthetic pathways, molecular breeding of varieties and provides medicinal plant resource information and data support for development and utilization of medicinal plants in China and abroad.

Prognostic value of serum free triiodothyronine in patients with hepatitis E-related acute liver failure / 中华临床感染病杂志

Objective:To investigate the prognostic value of serum free triiodothyronine (FT3) in patients with hepatitis E-related acute liver failure (HEV-ALF).Methods:Clinical data of 88 patients with HEV-ALF and 86 patients with acute hepatitis E (AHE) were collected from the member hospitals of Chinese Consortium for the Study of Hepatitis E between January 2016 and December 2021; the data of 100 health subjects who underwent health check-up in Suzhou Municipal Hospital were also collected as healthy control (HC) group. Serum FT3 levels were analyzed in all subjects. HEV-ALF patients were divided into survival group ( n=73) and death group ( n=15) according to their 30 day survival. Correlation between serum FT3 level and prognosis of HEV-ALF patients were analyzed by Cox regression and orthogonal partial least squares discriminant analysis (OPLS-DA). The receiver operating characteristic (ROC) curve and decision curve analysis (DCA) were used to assess the predictive value of serum FT3 levels for predicting the prognosis of patients, and its prediction efficacy was compared with conventional Model for End-Stage Liver Disease (MELD), King’s College Hospital criteria (KCH) and Child-Pugh models. Results:The levels of serum FT3 in HEV-ALF patients were significantly lower than those in AHE patients and HC group ( P=0.006 or <0.001). Cox regression analysis showed that international standardized ratio ( HR=17.984, 95% CI 2.804-115.362), hepatic encephalopathy ( HR=12.895, 95% CI 2.386-69.695) and total cholesterol ( HR=2.448, 95% CI 1.108-5.409) were independent risk factors for death in HEV-ALF patients, and serum FT3 level ( HR=0.323, 95% CI 0.119-0.876) was a protective factor. OPLS-DA results showed serum FT3 levels had high predictive value. ROC curve analysis results showed that the area under the curve was 0.828 (95% CI 0.733-0.900, P<0.001), the sensitivity was 80.00%, and the specificity was 78.08%. DCA showed that FT3 has good prediction ability and decision-making level serum FT3 levels in patients with improvement and fluctuation were significantly higher than those in the patients with deterioration ( P<0.05 or <0.01). Conclusion:Serum FT3 levels are closely related to the prognosis of HEV-ALF patients and it may be used as a biomarker for the prognosis of patients with HEV-ALF.

Corrigendum: TRPC1 Inhibits Cell Proliferation/Invasion and Is Predictive of a Better Prognosis of Esophageal Squamous Cell Carcinoma.

[This corrects the article DOI: 10.3389/fonc.2021.627713.].

Microtubule associated protein 4 (MAP4) phosphorylation reduces cardiac microvascular density through NLRP3-related pyroptosis.

Phosphorylation of MAP4 (p-MAP4) causes cardiac remodeling, with the cardiac microvascular endothelium being considered a vital mediator of this process. In the current study, we investigated the mechanism underlying p-MAP4 influences on cardiac microvascular density. We firstly confirmed elevated MAP4 phosphorylation in the myocardium of MAP4 knock-in (KI) mice. When compared with the corresponding control group, we detected the decreased expression of CD31, CD34, VEGFA, VEGFR2, ANG2, and TIE2 in the myocardium of MAP4 KI mice, accompanied by a reduced plasma concentration of VEGF. Moreover, we observed apoptosis and mitochondrial disruption in the cardiac microvascular endothelium of MAP4 KI animals. Consistently, we noted a decreased cardiac microvascular density, measured by CD31 and lectin staining, in MAP4 KI mice. To explore the underlying mechanism, we targeted the NLRP3-related pyroptosis and found increased expression of the corresponding proteins, including NLRP3, ASC, mature IL-1ß, IL-18, and GSDMD-N in the myocardium of MAP4 KI mice. Furthermore, we utilized a MAP4 (Glu) adenovirus to mimic cellular p-MAP4. After incubating HUVECs with MAP4 (Glu) adenovirus, the angiogenic ability was inhibited, and NLRP3-related pyroptosis were significantly activated. Moreover, both cytotoxicity and PI signal were upregulated by the MAP4 (Glu) adenovirus. Finally, NLRP3 inflammasome blockage alleviated the inhibited angiogenic ability induced by MAP4 (Glu) adenovirus. These results demonstrated that p-MAP4 reduced cardiac microvascular density by activating NLRP3-related pyroptosis in both young and aged mice. We thus managed to provide clues explaining MAP4 phosphorylation-induced cardiac remodeling and enriched current knowledge regarding the role of MAP4.

TRPC1 Inhibits Cell Proliferation/Invasion and Is Predictive of a Better Prognosis of Esophageal Squamous Cell Carcinoma.

BACKGROUND AND OBJECTIVES: In China, over 90% of esophageal cancer (EC) cases are esophageal squamous cell carcinoma (ESCC). ESCC is a frequently malignant tumor with poor prognosis despite the development of comprehensive therapeutic strategies, for which there is still a lack of effective prognostic factors. Previous studies found that the abnormal expression of TRPC1 is closely related to the proliferation, invasion, metastasis, and differentiation of various tumors. However, the relationship between TRPC1 and ESCC is currently unclear. The present study aimed to clarify the clinical significance of TRPC1 and to preliminarily assess the molecular mechanism by which TRPC1 regulates cell proliferation, migration, and invasion in ESCC. MATERIALS AND METHODS: Immunohistochemistry (IHC) was used to determine the expression of TRPC1 and Ki-67 in 165 cases of ESCC. The correlations between TRPC1 expression and clinicopathological characteristics were determined, and both univariate and multivariate analyses were utilized to quantify the impact of TRPC1 expression on patient survival. Cell Counting Kit-8, scratch wound healing, and transwell assays were used to determine the effects of TRPC1 on proliferation, migration, and invasion in ESCC in vitro, respectively. RESULTS: The positive expression rate of TRPC1 showed significantly decreased in ESCC (45.50%) compared with the levels in normal esophageal mucosa (NEM; 80.80%) and high-grade intraepithelial neoplasia (HGIEN; 63.20%) (P<0.001). Higher expression rate of TRPC1 was associated with low lymph node metastasis (P<0.001), high differentiation (rs = 0.232, P=0.003), and low Ki-67 (rs = -0.492, P<0.001). We further revealed that low expression of TRPC1 was associated with poor prognosis (Disease-free survival, DFS: 95% CI=0.545-0.845, P=0.001; Overall survival, OS: 95% CI=0.553-0.891, P=0.004). Furthermore, we showed that downregulation of TRPC1 promoted the proliferation, migration, and invasion of human esophageal squamous cell carcinoma cell line EC9706 in vitro. In contrast, overexpression of TRPC1 inhibited the proliferation, migration, and invasion of human esophageal squamous cell carcinoma cell line KYSE150 (P<0.01), in a manner at least in part mediated through the AKT/p27 pathway. CONCLUSION: TRPC1 inhibited the proliferation, migration, and invasion of EC9706 and KYSE150 cells, at least, in part mediated through the AKT/p27 pathway in vitro. The downregulation of TRPC1 may be one of the most important molecular events in the malignant progression of ESCC. TRPC1 could be a new candidate tumor suppressor gene and a new prognostic factor of ESCC.

TSH adenoma and syndrome of resistance to thyroid hormones-Two cases report of syndrome of inappropriate secretion of thyrotropin.

SITSH (syndrome of inappropriate secretion of thyrotropin) is a rare clinical state defined as uninhibited serum thyroid stimulating hormone in the presence of elevated thyroid hormone. This state is complicated and mainly caused by the abnormal feedback of hypothalamus-pituitary thyroid axis. The TSH adenoma (TSH-oma) and resistance to thyroid hormones (RTH) are the main etiologies of SITSH. As is well known that the treatment strategies of RTH and TSH-oma are apparently different, thus identifying the difference between RTH and TSH-oma is of great significance for the diagnosis and treatment of SITSH. CASE DESCRIPTION: A 62-year-old man with a state of elevated thyroid hormones and inappropriate elevated serum TSH level was hospitalized in 2016. Results of the pituitary enhanced magnetic resonance imaging and the somatostatin test respectively demonstrated a space-occupying lesion of pituitary and an elevated serum sex hormone binding globulin (SHBG) and inhibited TSH secretion, which indicated the occurrence of TSH-oma. In 2019, a 23-year-old girl with a state of elevated thyroid hormones and inappropriate normal serum TSH was hospitalized. Interestingly, whole exome sequencing detection suggested a pathogenic mutation in thyroid hormone receptor ß (THRB) gene, which has been shown to be associated with RTH. CONCLUSIONS: The difference between TSH-oma and RTH ought to be clarified for their accurate diagnose and treatment. The clinical experiences of the two cases reported here suggest that more detail information such as family medical history, serum SHBG level, and THRB gene test is helpful for the diagnose and treatment of TSH-oma and RTH. Additionally, we also summarized the identification points, diagnosis process, and treatment strategies for these two rare diseases.

Co-expression of VEGF-C and survivin predicts poor prognosis in esophageal squamous cell carcinoma.

BACKGROUND: Lymphatic metastasis is one of the main factors affecting prognosis in esophageal squamous cell carcinoma (ESCC). Vascular endothelial growth factor-C (VEGF-C) is an important factor that promotes lymphangiogenesis. Survivin also plays a significant role in lymphatic invasion. However, the role and mechanism of their co-expression are still unclear in ESCC. The purpose of this study was to investigate whether the co-expression of VEGF-C and survivin could be a potential marker to predict patient prognosis and survival in ESCC. METHODS: The levels of VEGF-C, vascular endothelial growth factor receptor 3 (VEGFR-3), survivin, and Ki-67 were determined by immunohistochemistry (IHC) in 97 ESCC patient tumors. The correlations of co-expression of VEGF-C and survivin with pathological features and survival results were also assessed. RESULTS: High VEGF-C expression was observed in 64.9% of the patients and significantly correlated with T stage (P=0.024), node status (P=0.038), and lymph node metastasis (P=0.015). High survivin expression was significantly associated with T stage (P=0.013), N stage (P=0.016), lymph node metastasis (P=0.005), and differentiation (P=0.044) in 67.0% of the patients. Co-expression of VEGF-C and survivin (V+S+) was significantly associated with T stage (P<0.001), N stage (P=0.015), lymph node metastasis (P=0.003), differentiation (P=0.0045), and Ki-67 levels (P=0.024). High expression of VEGF-C or survivin was associated significantly with worse disease-free survival (DFS) and overall survival (OS) (P<0.05). Moreover, the V+S+ group had a worse DFS (P<0.001) and OS (P=0.001) than any other group (i.e., V-S-, V+S-, V-S+). Furthermore, multivariate DFS analyses (95% CI: 1.147-2.220, P=0.006) and multivariate OS analyses (95% CI: 1.080-2.193, P=0.017) revealed that co-expression of VEGF-C and survivin was an independent prognostic factor in ESCC patients. CONCLUSIONS: Co-expression of VEGF-C and survivin was predictive of poor prognosis in ESCC. Combined detection of VEGF-C and survivin could represent a feasible and effective marker to predict the prognosis and survival of ESCC patients.

Comparison of Sonazoid contrast-enhanced ultrasound and contrast-enhanced MRI in the diagnosis of focal liver lesions / 中华超声影像学杂志

Objective:To compare the diagnostic efficacies of Sonazoid contrast-enhanced ultrasound (CEUS) and contrast-enhanced magnetic resonance imaging (CE-MRI) in the diagnosis of focal liver lesions (FLLs), and to evaluate the clinical value of Sonazoid.Methods:A total of 58 FLLs in 50 patients who underwent Sonazoid-CEUS and CE-MRI examinations from July 2019 to January 2021 in the First Affiliated Hospital of Zhejiang University School of Medicine were enrolled in this study according to the inclusion criteria. The final diagnostic reference standard was decided by surgical pathology or ultrasound-guided biopsy pathology. Sonazoid-CEUS and CE-MRI features of benign and malignant FLLs were analyzed, and the sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic coincidence rate of the two tests were calculated respectively.Results:There was a statistically significant difference between benign and malignant FLLs in the imaging pattern of homogeneous or heterogeneous intratumoral enhancement in the artery-dominant phase and washout images in the late phase( P<0.001).9.8%(4/41) of the malignant lesions did not decrease until the late phase but decreased in the post-vascular phase. The sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic coincidence rate of the two tests were 97.6%, 52.9%, 83.3%, 90.0%, 84.5%(Sonazoid-CEUS) and 85.4%, 64.7%, 85.4%, 64.7%, 79.3%(CE-MRI), the differences of sensitivity and specificity were not statistically significant ( P=0.125, P=0.687). Conclusions:The vascular phase in Sonazoid-CEUS is still an important diagnostic sign of FLLs, and the unique Kupffer phase can provide additional information for the diagnosis. Sonazoid-CEUS has the same important value as CE-MRI in the diagnosis of FLLs.

Clinical features and risk factors of invasive pulmonary aspergillosis in patients with chronic obstructive pulmonary disease / 中华临床感染病杂志

Objective:To analyze the clinical features and risk factors of invasive pulmonary aspergillosis (IPA) in patients with chronic obstructive pulmonary disease (COPD).Methods:Ninety COPD patients with IPA admitted in the First Affiliated Hospital, Zhejiang University School of Medicine and Zhejiang Provincial People’s Hospital from January 2015 to September 2019 were enrolled, and 180 COPD patients without IPA admitted in the same period were selected as control group. The clinical data of the patients in both groups were analyzed retrospectively. Chi square test was used to compare the imaging characteristics of patients in two groups, and multivariate conditional Logistic regression analysis was used to explore the risk factors of IPA in COPD patients. Results:Among 90 cases of COPD with IPA, the culture of lower respiratory tract samples identified Aspergillus fumigatus in 78 cases, Aspergillus flavus in 6 cases, Aspergillus fumigatus with Aspergillus flavus in 1 case, Aspergillu sterrus in 1 case, and Aspergillus nigerwere in 1 case; 1 case of Aspergillus mycelium was found by sputum exfoliation cytology and 2 cases were positive for serum galactomannan. Chest CT images showed patchy infiltrating shadow (87.8%), pleural effusion (36.7%), nodules (33.3%), cavity (18.9%), consolidation shadow (17.8%), halo sign (14.4%) and air crescent sign (2.2%). The incidence of patchy infiltrating shadow, consolidation shadow, halo sign and cavity were higher in COPD patients with IPA compared to control group ( P<0.05 or P<0.01). Multivariate conditional Logistic regression analysis showed that prolonged hospital stay ( OR=1.183, 95% CI 1.047-1.336), combination of two antibiotics ( OR=5.391, 95% CI 1.241-23.421), duration of antibiotic treatment ≥14 days ( OR=5.275, 95% CI 1.586-17.541), cumulative use of antibiotics ( OR=2.270, 95% CI 1.406-3.664) were the risk factors of COPD with IPA ( P<0.05 or P<0.01). Conclusion:The risk factors of IPA in COPD patients include long duration of hospital stay, combination of two kinds of antibiotics, more than 14 days of antibiotic treatment, and more varieties of antibiotics. If the above risk factors exist in patients with COPD, etiology and serology examination and dynamic monitoring of chest CT scan should be performed for early diagnosis and improved prognosis.

Population genetic diversity of an odorous frog Odorrana grahami (Amphibia: Anura: Ranidae) in relation to conservation based on mitochondrial DNA.

The diskless-fingered odorous frog Odorrana grahami is widely distributed in the high-altitude mountains in the southwestern China and northern Indochina regions. In this study, a comparative analysis of the mitochondrial COI gene sequences was performed to examine the population genetic diversity of 76 individuals from 10 localities across the distributional range of the species. Haplotype diversity and nucleotide diversity were 0.605 and 0.00199, respectively, in the total population. An AMOVA indicated that 47.3% of the total variation originated from variation within individual populations and 52.7% came from variation between the 10 geographic populations. Tests of neutral evolution indicated that a recent expansion occurred in total population. The findings provide useful information for the conservation of this species.

Chronic cadmium exposure aggravates malignant phenotypes of nasopharyngeal carcinoma by activating the Wnt/ß-catenin signaling pathway via hypermethylation of the casein kinase 1α promoter.

BACKGROUND: Our previous study has shown that cadmium (Cd) exposure is not only a risk factor for nasopharyngeal carcinoma (NPC), but also correlated with the clinical stage and lymph node metastasis. However, the underlying molecular events of Cd involved in NPC progression remain to be elucidated. PURPOSE: The objective of this study was to decipher how Cd impacts the malignant phenotypes of NPC cells. METHODS: NPC cell lines CNE-1 and CNE-2 were continuously exposed with 1 µM Cd chloride for 10 weeks, designating as chronic Cd treated NPC cells (CCT-NPC). MTT assay, colony formation assay and xenograft tumor growth were used to assess cell viability in vitro and in vivo. Transwell assays were performed to detect cell invasion and migration. The protein levels of E-cadherin, N-cadherin, Vimentin as well as ß-catenin and casein kinase 1α(CK1α) were measured by Western blot. Immunofluorescence staining was used to observe the distribution of filament actin (F-actin), ß-catenin and CK1α. The mRNA levels of downstream target genes of ß-catenin were detected by RT-PCR. Wnt/ß-catenin signaling activity was assessed by TOPFlash/FOPFlash dual luciferase report system. MS-PCR was used to detect the methylation status of CK1α. Finally, the activation of Wnt/ß-catenin pathway and cell biological properties were examined following treatment of CCT-NPC cells with 5-aza-2-deoxy-cytidine(5-aza-CdR). RESULTS: CCT-NPC cells showed an increase in cell proliferation, colony formation, invasion and migration compared to the parental cells. Cd also induced cytoskeleton reorganization and epithelial-to-mesenchymal transition. Upregulation and nuclear translocation of ß-catenin and increased luciferase activity accompanied with transcription of downstream target genes were found in CCT-NPC cells. Treatment of CCT-CNE1 cells with 5-aza-CdR could reverse the hypermethylation of CK1α and attenuate the cell malignancy. CONCLUSION: These results support a role for chronic Cd exposure as a driving force for the malignant progression of NPC via epigenetic activation of the Wnt/ß-catenin pathway.

Osthole sensitizes with radiotherapy to suppress tumorigenesis of human nasopharyngeal carcinoma in vitro and in vivo.

BACKGROUND: Radiotherapy is one of the most comment and useful treatment for nasopharyngeal carcinoma (NPC), but the radioresistance remains a major obstacle. Osthole, a natural coumarin derivative, has been shown to have anti-tumor and anti-inflammatory activity. However, the relationship between osthole and NPC treatment, especially for radiotherapy, is still elusive. METHODS: Osthole with or without X ray radiotherapy treated with CNE2 cells, a human EC cell line. Cell viability, proliferation, migration and apoptosis were measured by MTT, colony formation, Annexin V/PI double staining, Transwell assay, respectively. NPC tumor models were established on BALB/c nude mice by subcutaneously injection of CNE2 cells and the effect of osthole and radiotherapy on tumor growth in vivo was studied. RESULTS: We found that in a dose-dependent manner, osthole could individually, and synergistically with radiotherapy, reduce NPC cell (CNE2) viability, proliferation, migration, and invasion, and induce apoptosis, respectively. This effect of anti-tumor growth and induction of apoptosis was further confirmed in mice induced by subcutaneously injection with CNE2 cells and following treated with osthole or/and radiation. CONCLUSION: Osthole increases the effect of radiotherapy on anti-human nasopharyngeal cancer.

Effect of osthole on proliferation and radiosensitivity of poorly differentiated CNE2 stem cells from nasopharyngeal carcinoma / 中草药

Objective To investigate the effect and the underlying mechanism of osthole on the proliferation and radiosensitivity of CNE2 stem cells, one of the poorly differentiated cell lines from nasopharyngeal carcinoma. Methods Poorly differentiated CNE2 stem cells were isolated and cultured in serum-free medium (SFM). Flow cytometry was used to detect biomarkers (CD44+/CD24-low) of stem cells and the activity of ALDH. CNE2 stem cells was treated with different concentrations of osthole (0, 20, 40, 80 μg/mL), and then subject to MTT assay. Additionally, CNE2 stem cell was treated with 40 μg/mL osthole plus different dose of radiation (0, 2, 5 Gy) followed by colony formation assay. Consequently, Western blotting was used to detect the difference of pGSK-3β, β-catenin, and Cyclin D1 protein expression in CNE2 stem cells after the treatment of osthole with or without radiation. Results Poorly differentiated CNE2 stem cells isolated and cultured in serum-free medium (SFM) could be passaged stably. The ratio of CD44+/CD24-low and the activity of ALDH were significantly higher in CNE2 stem cells than that in the parental cells (P < 0.05). Compared to the control group, osthole could obviously suppress the proliferation of CNE2 stem cells in a dose and time dependent manner (P < 0.05). Moreover, colony formation assay revealed that inhibition rate of CNE2 stem cell colony formation was highest after the treatment of osthole plus radiation, followed by the treatment of osthole or radiation alone (P < 0.05). Western blotting indicated that in contrast to the controls, the expression of pGSK-3β, β-catenin, and Cyclin D1 protein was mostly down-regulated in the CNE2 stem cells treated with osthole plus radiation, followed by that treated with osthole or radiation alone (P < 0.05). Conclusion Osthole effectively inhibited the proliferation and increased the radiosensitivity of CNE2 stem cells, probably due to the down-regulation of pGSK-3, β-catenin, and Cyclin D1 in tumor stem cells by osthole.

The potential regulatory roles of NAD(+) and its metabolism in autophagy.

(Macro)autophagy mediates the bulk degradation of defective organelles, long-lived proteins and protein aggregates in lysosomes and plays a critical role in cellular and tissue homeostasis. Defective autophagy processes have been found to contribute to a variety of metabolic diseases. However, the regulatory mechanisms of autophagy are not fully understood. Increasing data indicate that nicotinamide adenine nucleotide (NAD(+)) homeostasis correlates intimately with autophagy. NAD(+) is a ubiquitous coenzyme that functions primarily as an electron carrier of oxidoreductase in multiple redox reactions. Both NAD(+) homeostasis and its metabolism are thought to play critical roles in regulating autophagy. In this review, we discuss how the regulation of NAD(+) and its metabolism can influence autophagy. We focus on the regulation of NAD(+)/NADH homeostasis and the effects of NAD(+) consumption by poly(ADP-ribose) (PAR) polymerase-1 (PARP-1), NAD(+)-dependent deacetylation by sirtuins and NAD(+) metabolites on autophagy processes and the underlying mechanisms. Future studies should provide more direct evidence for the regulation of autophagy processes by NAD(+). A better understanding of the critical roles of NAD(+) and its metabolites on autophagy will shed light on the complexity of autophagy regulation, which is essential for the discovery of new therapeutic tools for autophagy-related diseases.

Clinical importance and therapeutic implication of E-cadherin gene methylation in human ovarian cancer.

E-cadherin (E-cad) is widely expressed in epithelial cells and acts as a pivotal tumor suppressor. The promoter methylation of E-cad has been reported to closely relate to its downregulation in many kinds of cancers. E-cad expression and methylation status were detected by immunohistochemistry (IHC) and methylation-specific polymerase chain reaction (MS-PCR) in 50 ovarian cancer tissues. 5-Aza-2'-deoxycytidine (5-Aza-dC) was used to demethylate E-cad in SKOV3 and ES2 ovarian cancer cell lines, of which the effect was verified by Western blot and MS-PCR. Then MTT and transwell experiments were conducted to detect the capacity of cell proliferation and migration for these cells. Downregulation of E-cad expression was observed in 60 % of ovarian cancer tissues (30/50) by IHC, whereas MS-PCR result indicated that E-cad was methylated in 64 % of (32/50) ovarian cancer specimens. And E-cad expression was significantly correlated with E-cad methylation (P = 0.004). 5-Aza-dC was used to process SKOV3 and ES2 ovarian cancer cell lines. By MTT experiment, we found that the proliferation of 5-Aza-dC-treated SKOV3 and ES2 was significantly suppressed by 28.0 % (P < 0.05) and 32.3 % (P < 0.05). By transwell experiment, the motility of SKOV3 and ES2 was found to be significantly suppressed by 38.2 and 27.4 % (P < 0.05), respectively, after treated with 5-Aza-dC. E-cad methylation is one of the main reasons for the expression reduction in ovarian cancer. 5-Aza-dC treatment could significantly restore the expression of E-cad and suppress growth and invasion of SKOV3 and ES2 cells. These results suggest E-cad methylation may be a promising target for ovarian cancer therapy.

High-risk human papillomavirus infection associated with telomere elongation in patients with esophageal squamous cell carcinoma with poor prognosis.

BACKGROUND: Telomere maintenance is crucial in carcinogenesis and tumor progression. The results of a previous study from the authors indicated that infection with high-risk human papillomavirus (HR-HPV) types 16, 18, and 58 was a risk factor for esophageal squamous cell carcinoma (ESCC) in the Shantou region of China. In the current study, the authors explored the association between HR-HPV infection, telomere length (TL), and DNA methylation and their significance in the prognosis of patients with ESCC. METHODS: TL and DNA methylation were analyzed by real-time polymerase chain reaction and methylation-specific polymerase chain reaction in 70 cases of ESCC tumor (T) and paired nontumor (NT) tissues and 50 cases of normal esophagus (NE). The prognostic value of TL and DNA methylation in ESCC was analyzed. RESULTS: TL gradually decreased from NE to NT to T tissue. TL in tumor tissue (T-TL) was found to be longer in tissue that was positive for HR-HPV compared with negative tissue and was found to be positively associated with viral load (Spearman correlation, 0.410; P = .037) and integration (represented by the ratio of HR-HPV E2 to E6/E7 genes; P = .01). The DNA methylation ratio of human telomerase reverse transcriptase was more prevalent with long (≥ 0.7) compared with short (< 0.7) T-TL and was positively correlated with T-TL (Spearman correlation, 0.318; P = .007) and HR-HPV integration (P = .036). Furthermore, Cox proportional hazards modeling revealed a high ratio of T-TL to NT-TL (≥ 0.80) as a factor of poor prognosis, independent of other clinicopathologic variables. CONCLUSIONS: HR-HPV infection and integration related to telomere elongation and DNA methylation of human telomerase reverse transcriptase may be a potential biomarker of prognosis in patients with ESCC.

Identification of differentially expressed serum proteins in infectious purpura fulminans.

Purpura fulminans (PF) is a life-threatening hemorrhagic condition. Because of the rarity and randomness of the disease, no improvement in treatment has been made for a long time. In this study, we assessed the serum proteome response to PF by comparing serum proteins between healthy controls and PF patient. Liquid chromatography with tandem mass spectrometry (LC-MS/MS) approach was used after depleting 6 abundant proteins of serum. In total, 262 proteins were confidently identified with 2 unique peptides, and 38 proteins were identified significantly up- (≥ 2) or downregulated (≤ 0.5) based on spectral counting ratios (SpCPF/N). In the 38 proteins with significant abundance changes, 11 proteins were previously known to be associated with burn or sepsis response, but 27 potentially novel proteins may be specifically associated with PF process. Two differentially expressed proteins, alpha-1-antitrypsin (SERPINA1) and alpha-2 antiplasmin (SERPINF2), were validated by Western blot. This is the first study where PF patient and healthy controls are compared in a proteomic study to elucidate proteins involved in the response to PF. This study provides an initial basis for future studies of PF, and the differentially expressed proteins might provide new therapeutic targets to decrease the mortality of PF.

Follow-up study of 1035 children with recurrent wheezing / 临床儿科杂志

Objective To investigate the long-term outcome of children with recurrent wheeze and to determine the effectiveness of inhaled hormone therapy. Methods One thousand and thirty-five children with recurrent wheezing were followed up for more than 4 years and the data were retrospectively evaluated. Results Of 1035 cases, 751 (72.56%) patients outgrew their wheeze during the follow-up period, whereas the other 284 (27.44%) patients had recurrence wheeze during the last two years. The age of wheezing onset was<3 years in 542 (52.37%) cases, from 3 to 7 years in 386 (37.29%) cases, and from 7 to 12 years in 107 (10.34%) cases. There was significant difference in clinical control rate among groups with different wheezing ages onset (χ2=45.27, P<0.001). Children with wheezing age onset from 7 to 12 years had the lowest clinical control rate. Among 1035 wheeze children, 343 (79.95%) children in 429 cases who received inhaled hormone therapy for more than one year outgrew their wheeze. Whereas 408 (67.35%) in 606 cases who did not receive inhaled hormone therapy outgrew their wheeze. There was significant difference of clinical control rate between inhaled group and non-inhaled group (P<0.01). Con-clusions The age of wheezing onset is<7 years in 89.66%of children with recurrent wheeze. Most of them can be clinicalycon-trolled. The long term inhaled hoemone therapy for children with recurrent wheeze can reduce the risk of developing adulthood asthma.

Phosphorylation of DYNLT1 at serine 82 regulates microtubule stability and mitochondrial permeabilization in hypoxia.

Hypoxia-induced microtubule disruption and mitochondrial permeability transition (mPT) are crucial events leading to fatal cell damage and recent studies showed that microtubules (MTs) are involved in the modulation of mitochondrial function. Dynein light chain Tctex-type 1 (DYNLT1) is thought to be associated with MTs and mitochondria. Previously we demonstrated that DYNLT1 knockdown aggravates hypoxia-induced mitochondrial permeabilization, which indicates a role of DYNLT1 in hypoxic cytoprotection. But the underlying regulatory mechanism of DYNLT1 remains illusive. Here we aimed to investigate the phosphorylation alteration of DYNLT1 at serine 82 (S82) in hypoxia (1% O2). We therefore constructed recombinant adenoviruses to generate S82E and S82A mutants, used to transfect H9c2 and HeLa cell lines. Development of hypoxia-induced mPT (MMP examining, Cyt c release and mPT pore opening assay), hypoxic energy metabolism (cellular viability and ATP quantification), and stability of MTs were examined. Our results showed that phosph-S82 (S82-P) expression was increased in early hypoxia; S82E mutation (phosphomimic) aggravated mitochondrial damage, elevated the free tubulin in cytoplasm and decreased the cellular viability; S82A mutation (dephosphomimic) seemed to diminish the hypoxia-induced injury. These data suggest that DYNLT1 phosphorylation at S82 is involved in MTs and mitochondria regulation, and their interaction and cooperation contribute to the cellular hypoxic tolerance. Thus, we provide new insights into a DYNLT1 mechanism in stabilizing MTs and mitochondria, and propose a potential therapeutic target for hypoxia cytoprotective studies.

miR-27b represses migration of mouse MSCs to burned margins and prolongs wound repair through silencing SDF-1a.

BACKGROUND: Interactions between stromal cell-derived factor-1α (SDF-1α) and its cognate receptor CXCR4 are crucial for the recruitment of mesenchymal stem cells (MSCs) from bone marrow (BM) reservoirs to damaged tissues for repair during alarm situations. MicroRNAs are differentially expressed in stem cell niches, suggesting a specialized role in stem cell regulation. Here, we gain insight into the molecular mechanisms involved in regulating SDF-1α. METHODS: MSCs from green fluorescent protein transgenic male mice were transfused to irradiated recipient female C57BL/6 mice, and skin burn model of bone marrow-chimeric mice were constructed. Six miRNAs with differential expression in burned murine skin tissue compared to normal skin tissue were identified using microarrays and bioinformatics. The expression of miR-27b and SDF-1α was examined in burned murine skin tissue using quantitative real-time PCR (qPCR) and immunohistochemistry (IHC), enzyme-linked immunosorbent assay (ELISA). The Correlation of miR-27b and SDF-1α expression was analyzed by Pearson analysis Correlation. miRNAs suppressed SDF-1α protein expression by binding directly to its 3'UTR using western blot and luciferase reporter assay. The importance of miRNAs in MSCs chemotaxis was further estimated by decreasing SDF-1α in vivo and in vitro. RESULTS: miR-23a, miR-27a and miR-27b expression was significantly lower in the burned skin than in the normal skin (p<0.05). We also found that several miRNAs suppressed SDF-1α protein expression, while just miR-27a and miR-27b directly bound to the SDF-1α 3'UTR. Moreover, the forced over-expression of miR-27a and miR-27b significantly reduced the directional migration of mMSCs in vitro. However, only miR-27b in burn wound margins significantly inhibited the mobilization of MSCs to the epidermis. CONCLUSION: miR-27b may be a unique signature of the stem cell niche in burned mouse skin and can suppress the directional migration of mMSCs by targeting SDF-1α by binding directly to its 3'UTR.