[Call for attention to the value and challenge of liquid biopsy in diagnosis and treatment for peritoneal metastasis of gastric cancer].

Peritoneal metastasis is the common route of metastasis in gastric cancer and is a major cause of death in advanced gastric cancer. Early intervention with comprehensive treatment can effectively improve the prognosis of some patients with peritoneal metastasis. However, early peritoneal metastasis in gastric cancer is predominantly micro-metastasis, which cannot be effectively evaluated by imaging studies. Moreover, the detection of disseminated cancer cells in peritoneal lavage suffers from a low detection rate and significant heterogeneity. In recent years, the development and application of new liquid biopsy technologies such as circulating tumor cells (CTCs) and circulating tumor DNA (ctDNA) have provided new means to assess potential peritoneal metastasis at the cellular and molecular levels, gradually becoming research hotspots in this field. This review will summarize the relevant progress of liquid biopsy in peritoneal metastasis, which holds significant importance for improving the prognosis of gastric cancer patients in China.

[Association between body mass index and coronary heart disease in Qingdao: a prospective study].

Objective: To analyze the association between body mass index (BMI) and coronary heart disease. Methods: The data for the present study were from the prospective cohort study of China Kadoorie Biobank (CKB) in Qingdao, a total of 33 355 participants aged 30-79 years were included in the study. Cox regression analyses were performed to evaluate the association between BMI and coronary heart disease. Results: During the follow-up for an average 9.2 years, a total of 2 712 cases of ischemic heart disease (IHD) and 420 cases of major coronary events (MCE) were found. Multivariate Cox regression analysis showed that, compared with participants with normal BMI, the participants who were overweight had a 41% and 87% higher risk of IHD and MCE, the adjusted HR were 1.41 (95%CI: 1.27-1.56) and 1.87 (95%CI: 1.43-2.44), respectively. The participants who were obesity had 91% and 143% higher risk of IHD and MCE, the adjusted HR were 1.91 (95%CI: 1.72-2.13) and 2.43 (95%CI: 1.82-3.24), respectively. Conclusion: Overweight and obesity might increase the risk for IHD and MCE.

[Standardized implementation of minimally invasive surgery for Siewert â ¡ esophagogastric junction adenocarcinoma].

The incidence of Siewert type II adenocarcinoma of the esophagogastric junction (AEG) is increasing year by year. Due to its special anatomical location and biological behavior, the treatment of AEG is still controversial in terms of lymph node dissection, the esophageal resection margin, range of gastrectomy, and the choice of reconstruction modality for postoperative gastrointestinal tract. The advent of the minimally invasive era has brought the treatment of Siewert type II AEG to a stage of gradual improvement and standardization. Experts of China are also actively exploring the value of minimally invasive surgery in the treatment of AEG through multicenter trials (CLASS-10, etc.). It is believed that based on the active development of many clinical studies, basic experimental studies and large prospective clinical studies, the strengthening of communication and cooperation among various disciplines and the innovative application of new technologies can bring greater survival benefits to patients.

Prognostic significance of palliative gastrectomy in incurable advanced gastric cancer: a retrospective cohort study and meta-analysis.

OBJECTIVE: There is controversy regarding the role of palliative gastrectomy in patients with incurable advanced gastric cancer requiring surgical intervention. The present retrospective cohort study and meta-analysis aimed to determine whether palliative gastrectomy plus chemotherapy can prolong the survival of patients with incurable advanced gastric cancer requiring surgical intervention. PATIENTS AND METHODS: The data from 153 patients diagnosed with incurable advanced gastric cancer requiring surgical intervention at our institute between January 2000 and December 2012 were retrospectively reviewed. We analyzed the value of palliative gastrectomy and identified the potential prognostic factors. We also conducted a meta-analysis of 10 studies to validate our results. RESULTS: Multivariate analysis indicated that palliative gastrectomy was a favorable independent prognostic factor for prolonged overall survival in incurable advanced gastric cancer patients requiring surgical intervention (p=0.029). The median survival of patients who underwent palliative gastrectomy plus chemotherapy was significantly longer than that of those who underwent non-resection surgery plus chemotherapy (12 months vs. 9 months, p=0.020). The patients in the non-resection surgery plus chemotherapy group exhibited significantly shorter overall survival than those in the D1+ lymphadenectomy group, D2 lymphadenectomy group, or distal gastrectomy group (p=0.021, p=0.007, and p=0.006, respectively). Our meta-analysis revealed that gastrectomy plus chemotherapy improved long-term survival in incurable advanced gastric cancer patients (hazard ratio (HR): 0.48; 95% confidence interval (CI): 0.35-0.67; p<0.001). CONCLUSIONS: Palliative gastrectomy plus chemotherapy may improve overall patient survival compared with non-resection operations plus chemotherapy in incurable advanced gastric cancer patients requiring surgical intervention.

Long noncoding RNA CASC2 inhibits metastasis and epithelial to mesenchymal transition of lung adenocarcinoma via suppressing SOX4.

Since this article has been suspected of research misconduct and the corresponding authors did not respond to our request to prove originality of data and figures, "Long noncoding RNA CASC2 inhibits metastasis and epithelial to mesenchymal transition of lung adenocarcinoma via suppressing SOX4, by D. Wang, Z.-M. Gao, L.-G. Han, F. Xu, K. Liu, Y. Shen, published in Eur Rev Med Pharmacol Sci 2017; 21 (20): 4584-4590-PMID: 29131257" has been withdrawn. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/13630.

Role of sonoelastography in assessment of axillary lymph nodes in breast cancer: a systematic review and meta-analysis.

AIM: To evaluate the effectiveness of shear-wave elastography (SWE) and strain elastography (SE) for axillary lymph nodes (ALNs). MATERIALS AND METHODS: PubMed, Embase, and Cochrane Library databases were searched until September 2018. Weighted mean difference was calculated for continuous variables. The accuracy of sonoelastography was assessed by calculating pooled sensitivity, specificity, area under the curve (AUC), positive likelihood ratio (PLR), negative likelihood ratio (NLR), and diagnostic odds ratio (DOR). All data were analysed using Stata 12.0. RESULTS: Ten studies with 1,038 ALNs were included in the meta-analysis. Five studies evaluated the use of SE, and the other five evaluated the SWE. The SWE stiffness values of malignant ALNs were significantly higher than those of benign nodes. Both SE and SWE have relatively high specificity and sensitivity. The max stiffness in SWE showed the highest specificity (0.94; 95% confidence interval [CI], 0.81-0.98), PLR (12.1; 95% CI, 4-36.5), NLR (0.29; 95% CI, 0.12-0.69), AUC (0.94; 95% CI, 0.91-0.96), and DOR (42; 95% CI, 12-154); in contrast, the mean stiffness showed the highest sensitivity (0.80; 95% CI, 0.61-0.91). CONCLUSION: Sonoelastography demonstrated high sensitivity and specificity for differentiating between malignant and benign ALNs. The max and mean stiffness on SWE appeared to exhibit the highest accuracy. Thus, SWE is an effective accompaniment to sentinel node biopsy, and is appropriate for preoperative assessment of ALNs in the post-Z0011 era.

Comparison of Billroth I, Billroth II, and Roux-en-Y reconstructions after distal gastrectomy according to functional recovery: a meta-analysis.

OBJECTIVE: Gastric cancer is common, with a high mortality rate. Billroth I (B-I), Billroth II (B-II), and Roux-en-Y (R-Y) are the major reconstruction procedures after distal gastrectomy. In our study, we aimed to evaluate the functional recovery following the B-I, B-II, and R-Y reconstructions through a network meta-analysis. MATERIALS AND METHODS: PubMed, Embase, and Cochrane Library databases were searched until April 2018. From the included studies, first oral-intake time, early complications, endoscopic finding, quality of life (QoL), and body weight changes were extracted as the short- and long-term outcomes of reconstructions. The network meta-analysis was performed with R 3.4.2 software as well as "gemtc" and "forestplot" packages. RESULTS: Our work included a total of 26 articles involving 6212 patients with gastric cancer. Network meta-analysis revealed that R-Y reconstruction has a lower risk and degree of residual gastritis and bile reflex than B-I and B-II reconstructions. However, no differences in first oral-intake time, complications, risk of reflux esophagitis, and residual food, QoL, and body weight changes existed among the three reconstructions. CONCLUSIONS: R-Y may be the appropriate reconstruction procedure after distal gastrectomy based on postoperative functional recovery. However, more reports with a large sample size are warranted to investigate its long-term outcomes.

LncRNA TUC338 promotes invasion of lung cancer by activating MAPK pathway.

OBJECTIVE: Lung cancer is highly heterogeneous and the 5-year survival rate is less than 15%. It is currently difficult to determine the heterogeneity of lung cancer and the underlying pathogenetic of metastasis. We aimed to investigate the effect of long non-coding RNA (lncRNA) TUC338 on the invasion of lung cancer by activating MAPK pathway and to understand the heterogeneity and metastasis mechanism of lung cancer. PATIENTS AND METHODS: The expression of lncRNA TUC338 in 42 samples of lung cancer and paracancerous tissues were accessed by RT-qPCR. The relationship between the expression of lncRNA and clinicopathological parameters was analyzed. After overexpressing and interfering with lncRNA TUC338, effects of lncRNA TUC338 on cell proliferation and invasion were determined by cell counting kit-8 (CCK8) and transwell assay. The protein expression was evaluated by Western blot. RESULTS: Higher expression of TUC338 in lung cancer was observed in comparison with that in paracancerous tissues. The survival time of TUC338 was correlated with the expression of TUC338. Clinical data analysis revealed that the expression of TUC338 was correlated with the overall survival, tumor size and lymph node metastasis in patients, but not with age and gender. After interfering and overexpressing TUC338, it was found that the activity of lung cancer cells was decreased, as well as the invasion ability after interference with TUC338. After overexpression of TUC338, we found that lung cancer cell activity increased, as well as the invasion ability. By Western blot, we found that TUC338 can promote the development of lung cancer through regulating MAPK pathway. CONCLUSIONS: TUC338 was overexpressed in lung cancer, and its expression may have a relationship to the prognosis of lung cancer. MAPK pathway was involved in the invasion of lung cancer regulated by TUC338.

Long noncoding RNA CASC2 inhibits metastasis and epithelial to mesenchymal transition of lung adenocarcinoma via suppressing SOX4.

OBJECTIVE: Recently, long non-coding RNAs (lncRNAs) have caught more attention for their role in tumor progression. Lung adenocarcinoma (LAC) is one of these ordinary malignant tumors. This study aims to identify whether lncRNA CASC2 (cancer susceptibility candidate 2) can regulate the metastasis of LAC, and find out its potential mechanism. PATIENTS AND METHODS: RT-qPCR was conducted to detect CASC2 expression level in 63 LAC tissues and 4 LAC cells. Besides, statistical methods were applied to analyze clinical data and prognosis in the 63 patients. Furthermore, function experiments were performed to determine the effect of CASC2 on LAC metastasis in vitro. The potential mechanism was further explored by RT-qPCR and Western blot assay. RESULTS: In our study, CASC2 expression level was lower in LAC tissues than that in corresponding tissues. CASC2 expression was associated with lymph node metastasis, clinical stage and survival time of these patients. Moreover, overexpression of CASC2 inhibited migrated and invaded ability of LAC cells. Then, epithelial to mesenchymal transition (EMT) process of LAC cells and SOX4 expression was suppressed by upregulating CASC2. CONCLUSIONS: These results indicate that CASC2 could inhibit metastasis and EMT of LAC via suppressing SOX4, which may offer a new vision for interpreting the mechanism of LAC metastasis.

Caffeic acid n-butyl ester-triggered necrosis-like cell death in lung cancer cell line A549 is prompted by ROS mediated alterations in mitochondrial membrane potential.

OBJECTIVE: The aim of the present study was to evaluate the anticancer activity of caffeic acid n-butyl ester against lung cancer cell line A549 and to investigate the underlying mechanism. MATERIALS AND METHODS: IC50 was determined by MTT assay. Fluorescent probes DCFH-DA, Indo 1/AM, DiOC6 were used to determine ROS, Ca2+, and mitochondrial membrane potential (ΔΨm). ATP levels were determined by using ATP liteTM kit. DNA damage was investigated by DAPI and comet assays. Protein expression was investigated by Western blotting. RESULTS: Caffeic acid n-butyl ester exhibited lowest IC50 of 25 µM against lung A549 cell line. Caffeic acid n-butyl ester reduced the cell viability of A549 cells concentration and time-dependently. It also augmented the discharge of ROS and Ca2+ and lessened the mitochondrial membrane potential (ΔΨm) and ATP levels in A549 cells. Additionally, caffeic acid n-butyl ester also prompted DNA damage in A549 cell line. Notably, caffeic acid n-butyl ester-stimulated the cytochrome c release only and exhibited no effect on the expression of apoptosis-related protein levels such as caspase-3, caspase-8, and Apaf-1. DISCUSSION: Caffeic acid n-butyl ester exhibited significant anticancer activity against lung cancer cell line A549. However, the anticancer activity was not due to apoptosis as no significant change was observed in the expression of apoptosis-related proteins. The anticancer activity of caffeic acid n-butyl ester may be attributed to necrosis-like cell death prompted by ROS-mediated alterations in ΔΨm. CONCLUSIONS: Taken together, we conclude that caffeic acid n-butyl ester-induced A549 cells death displayed a cellular pattern characteristic of necrotic cell death and not of apoptosis.

Research on rat models of hypobaric hypoxia-induced pulmonary hypertension.

OBJECTIVE: Rat models of hypobaric hypoxia-induced pulmonary hypertension are commonly used in studies of chronic mountain sickness, while there are few researches specially focusing on these rats model. This study aims to exploring possible pathogenesis of hypobaric hypoxia-induced pulmonary hypertension by experimenting on hypobaric hypoxia-induced PH rat models at different simulate- altitudes. MATERIALS AND METHODS: 32 healthy male SD rats were randomly divided into six groups of different degree and time period of hypobaric hypoxia. The mean pulmonary arterial pressure (m PAP), right ventricular pressure (RVSP), the right ventricle (RV), left ventricular (LV), ventricular septal (S), the right ventricular hypertrophy index (RVHI) [calculated under the formula of RV / (LV + S)], hematoxylin-eosin staining, elastic fibers staining, the ratio of the thickness of vascular wall to its outer diameter (MT%), the ratio of the cross-sectional area of the middle vascular wall to the total vascular cross-sectional area (MA%); the α-SMA, and the Ki6 expressions were detected to evaluated the pulmonary hypertension. RESULTS: There were significant differences of the mPAP, RVSP and RVHI value between the hypobaric hypoxia groups and the control group (p < 0.05). The mPAP, RVSP, RVHI, MT%, MA%, α-SMA, and Ki6 of rats in model groups at an altitude of 3KM were higher than those of the control group, which raised gradually with the number of weeks increasing. The mPAP, RVSP, RV / (LV + S) value, MT%, MA%, α-SMA, and Ki67 of the 5KM-4W group were significantly higher than those of the control group (p < 0.05). CONCLUSIONS: Rat models with pulmonary hypertension at different altitudes have been successfully established by automatic adjusting hypobaric hypoxia chamber. Exposure to a low oxygen environment at a simulate-altitude of 3 km for 8 weeks have caused the pathological remodeling of pulmonary vascular walls and pulmonary hypertension, and further led to a series of pathological changes, including right ventricular hypertrophy. This model is easy to be replicated with good reproducibility and provides evidence for clinical trial of drugs.

Role of the NO/cGMP pathway in postoperative vasodilation in perforator flaps.

BACKGROUND: The nitric oxide (NO)/cyclic guanylyl monophosphate (cGMP) pathway is one of the most important regulators of tissue perfusion. Here, we sought to elucidate the protective effects of the NO/cGMP pathway on the microcirculation of axial pattern skin flaps. MATERIAL AND METHODS: Overall 40 rats were divided into four groups (n = 10 each): group A, sildenafil was administered orally at 10 mg/kg daily; group B, sildenafil citrate (10 mg/kg, oral) and nitro-amino-methyl-L-arginine (L-NAME, intraperitoneal injection), a nitric oxide synthase inhibitor, were administered daily; group C, L-NAME was administered alone; and group D, no drugs were administered. After surgery, the surviving flap area was calculated as a percentage of total flap dimensions using the paper template technique. Angiography and imaging were performed to compare the macrovascular changes of the choke zones in the flaps. Histological examinations were performed to compare the differences in microvascular changes between the two choke zones. RESULTS: A significant improvement of flap survival area and a significant dilation of vessels in both choke zones were found after administration of sildenafil. We also found that the postoperative vasodilation of choke vessels could be altered by inhibition of NO synthase (NOS). Moreover, the vasodilatory effect prolonged by the phosphodiesterase 5 inhibitor sildenafil was attenuated after administration of L-NAME. L-NAME significantly reversed the protection afforded by sildenafil. CONCLUSIONS: Targeting the NO/cGMP pathway can dilate vessels along the axis of the flap, including the choke vessels, thus augmenting flap viability. Therefore, targeting of this pathway may have therapeutic applications.

Genetic diversity and pathogenicity differentiation of Sclerotinia sclerotiorum on rapeseed (Brassica napus L.) in Anhui Province, China.

The pathogenicity of 47 isolates of Sclerotinia sclerotiorum from oilseed rape (Brassica napus L.) in Anhui, China, was tested by detached leaf inoculation using the susceptible rape cultivar Wanyou-14. All isolates were pathogenic to the cultivar and could be grouped into 3 categories based on the lesion length on the leaves tested: weak pathogenicity type, intermediate pathogenicity type, and strong pathogenicity type. This suggested that there was differentiation in the pathogenicity among the strains tested of S. sclerotiorum. Additionally, the intraspecific DNA polymorphisms among 47 strains of S. sclerotiorum were investigated by screening 40 pairs of inter-simple sequence repeat (ISSR) primers. Unweighted pair-group method with arithmetic average cluster analysis of these ISSR data distinguished all strains from each other and revealed considerable genetic variability among them. These strains were classified into 7 clusters according to their branching in the dendrogram, and partial correlation was observed between the genetic polymorphisms and the pathogenicity of S. sclerotiorum strains.

Treatment of seawater immersion-complicated open-knee joint fracture.

The current study aimed to select suitable remedies for seawater immersion-complicated open-knee joint fracture by exploring the effects of different treatment methods. Forty adult rabbits weighing 2.20 ± 0.25 kg were divided equally into internal fracture fixation group (A), seawater-immersed group with primary internal fixation (B), seawater-immersed group with secondary internal fixation (C), and seawater-immersed group with external fixation (D), using the random-digit table method. Open-femoral internal condylar fracture models were established. Group A was left untreated for 2 h, whereas the other three groups were subjected to seawater immersion for 2 h. Afterwards, groups A and B underwent debridement and steel plate and screw internal fixation. Group C underwent debridement and external fixation, which was followed by secondary steel plate and screw internal fixation after the wound healed. Group D underwent transarticular arthrodesis. Wound infection, joint functional rehabilitation, and radiological and histopathological changes in fracture healing in each group were assessed. The results showed that delayed internal fixation effectively reduces the infection rate of seawater immersion-complicated open fracture and benefits joint function rehabilitation.

First Report of Brown Blight Disease Caused by Colletotrichum gloeosporioides on Camellia sinensis in Anhui Province, China.

Yellow Mountain fuzz tip, a cultivar of Camellia sinensis (L.) Kuntze, is commonly grown in the Yellow Mountain region in Anhui Province of China. During 2011 to 2012, leaf and twig blight on tea plants occurred from July to September in growing regions. Symptoms of blight on leaves of infected plants were detected in 30 to 60% of the fields visited and up to 500 ha were affected each year. Symptoms began as small, water-soaked lesions on young leaves and twigs and later became larger, dark brown, necrotic lesions, 1 to 3 mm in diameter on leaves and 2 to 5 mm long on twigs. To determine the causal agent, symptomatic leaf tissue was collected from plants in Gantang and Tangkou townships in September 2012. Small pieces of diseased tea leaves and twigs were surface-disinfested in 2% NaClO for 3 min, rinsed twice in distilled water, plated on potato dextrose agar, and incubated at 28°C for 5 days. Eleven isolates were recovered and all cultures produced white-to-gray fluffy aerial hyphae and were dark on the reverse of the plate. The hyphae were hyaline, branching, and septate. Setae were 2- to 3-septate, dark brown, acicular, and 78.0 to 115.0 µm. Conidiogenous cells were hyaline, short, branchless, cylindrical, and 11.3 to 21.5 × 4.2 to 5.3 µm. Conidia were hyaline, aseptate, guttulate, cylindrical, and 12.5 to 17.3 × 3.9 to 5.8 µm. Appresoria were ovate to obovate, dark brown, and 8.4 to 15.2 × 7.8 to 12.9 µm. DNA was amplified using the rDNA-ITS primer pair ITS4/ITS5 (3), glyceraldehyde-3-phosphate dehydrogenase gene (GAPDH) primer pair GDF/GDR (2) and beta-tubulin 2 gene (Tub2) primer pair Btub2Fd/Btub4Rd (4). Sequences (GenBank Accession Nos. KC913203, KC913204, and KC913205) of the 11 isolates were identical and revealed 100% similarity to the ITS sequence of strain P042 of Colletotrichum gloeosporioides (EF423527), 100% identity to the GAPDH of isolate C07009 of C. gloeosporioides (GU935860), and 99% similarity to Tub2 of isolate 85 of C. gloeosporioides (AJ409292), respectively. Based on the above data, the 11 isolates were identified as C. gloeosporioides (Penz.) Penz. & Sacc. To confirm pathogenicity, Koch's postulate was performed and 4 ml of conidial suspension (1 × 105 conidia/ml) of each of the 11 isolates was sprayed on five leaves and five twigs per plant on four 12-month-old Yellow Mountain fuzz tip plants. Control plants were sprayed with distilled water. The inoculated plants were maintained at 28°C in a greenhouse with constant relative humidity of 90% and a 12-h photoperiod of fluorescent light. Brown necrotic lesions appeared on leaves and twigs after 7 days, while the control plants remained healthy. The experiments were conducted three times and the fungus was recovered and identified as C. gloeosporioides by both morphology and molecular characteristics. Tea plant blight caused by C. gloeosporioides was identified in Brazil (1), but to our knowledge, this is the first report of C. gloeosporioides causing tea leaf and twig blight on Yellow Mountain fuzz tip plants in Anhui Province of China. References: (1) M. A. S. Mendes et al. Page 555 in: Embrapa-SPI/Embrapa-Cenargen, Brasilia, 1998. (2) M. D. Templeton et al. Gene 122:225, 1992. (3) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990. (4) J. H. C. Woudenberg et al. Persoonia 22:56, 2009.

First Report of Leaf Spot Caused by Colletotrichum spaethianum on Peucedanum praeruptorum in China.

Peucedanum praeruptorum Dunn, a traditional Chinese medicinal herb, is an important crop in Ningguo, China. Since 2010, leaf spot symptoms were observed yearly starting in June. Blighted leaf areas on individual plants ranged from 10 to 25% in many fields, and up to 200 ha were affected each year. Symptoms consisted of small, brown, necrotic spots uniformly distributed on the 1- to 2-week-old leaves. Small tissue pieces from the edges of lesions were disinfected in 2% NaClO for 3 min, rinsed twice in distilled water, plated on potato dextrose agar (PDA), and incubated at 25°C in darkness for 4 days. Single spore isolations were obtained for six strains. When inoculated on SNA media, the six strains produced typical septate mycelium, with the young hyphae hyaline and aged ones white greyish. Setae of the strains on SNA were brown, tip acute, 2- to 3-septate, and 32.5 to 85.6 µm long. Conidiogenous cells were hyaline, cylindrical, 2- to 3-septate, 6.2 to 16.5 µm in length, and 2.8 to 4.3 µm in width. The mature conidia were slightly curved, with round apex and truncate base, 1 to 5 oil globules, and were 13.3 to 23.8 µm in length and 3.0 to 3.9 µm in width, respectively. Appressoria were solitary or in loose groups, dark brown, irregular shapes, and were 6.8 to 9.2 µm in length and 4.3 to 7.1 µm in width. PCR amplification was carried out by utilizing the universal rDNA-ITS primer pair ITS4/ITS5 (1) and the actin gene primer pair ACT-512F and ACT-783R (2). The PCR products of ITS (GenBank Accession No. KC913201) and actin gene (KC913202) from six isolates were identical, respectively, and shared 100% identity to the ITS sequence of strain CBS 167.49 of Colletotrichum spaethianum (GU227807.1) and 99% similarity to the actin gene of strain CBS 167.49 of C. spaethianum (GU227905.1), which was isolated from Hosta sieboldiana in Germany (3). Based on the above, the isolates were identified as C. spaethianum. To confirm pathogenicity, conidial suspensions (105 conidia ml-1) of each of the six isolates were sprayed on four leaves per plant on five 6-month-old P. praeruptorum plants. Control plants were sprayed with water. Plants were maintained at 28°C in a greenhouse with constant humidity (RH 90%) and a 12-h photoperiod of fluorescent light. Symptoms similar to the original ones started to appear after 10 days, while the control plants remained healthy. The tests were repeated three times and the fungus was recovered and identified as C. spaethianum by both morphology and molecular characterization. To our knowledge, this is the first report of C. spaethianum causing leaf spot on P. praeruptorum in China. Since the C. spaethianum infections pose a serious threat to P. praeruptorum production, this disease needs to be considered for developing effective control strategies. References: (1) I. Carbone and L. M. Kohn. Mycologia 91:553, 1999. (2) U. Damm et al. Fung. Divers. 39:45, 2009. (3) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.

Characterization of the interaction between astrocytes and encephalitogenic lymphocytes during the development of experimental autoimmune encephalitomyelitis (EAE) in mice.

The nature of pathogenic mechanisms associated with the development of multiple sclerosis (MS) have long been debated. However, limited research was conducted to define the interplay between infiltrating lymphocytes and resident cells of the central nervous system (CNS). Data presented in this report describe a novel role for astrocyte-mediated alterations to myelin oligodendrocyte glycoprotein (MOG)(35-55) -specific lymphocyte responses, elicited during the development of experimental autoimmune encephalitomyelitis (EAE). In-vitro studies demonstrated that astrocytes inhibited the proliferation and interferon (IFN)-γ, interleukin (IL)-4, IL-17 and transforming growth factor (TGF)-ß secretion levels of MOG(35-55) -specific lymphocytes, an effect that could be ameliorated by astrocyte IL-27 neutralization. However, when astrocytes were pretreated with IFN-γ, they could promote the proliferation and secretion levels of MOG(35-55) -specific lymphocytes, coinciding with apparent expression of major histocompatibility complex (MHC)-II on astrocytes themselves. Quantitative polymerase chain reaction (qPCR) demonstrated that production of IL-27 in the spinal cord was at its highest during the initial phases. Conversely, production of IFN-γ in the spinal cord was highest during the peak phase. Quantitative analysis of MHC-II expression in the spinal cord showed that there was a positive correlation between MHC-II expression and IFN-γ production. In addition, astrocyte MHC-II expression levels correlated positively with IFN-γ production in the spinal cord. These findings suggested that astrocytes might function as both inhibitors and promoters of EAE. Astrocytes prevented MOG(35-55) -specific lymphocyte function by secreting IL-27 during the initial phases of EAE. Then, in the presence of higher IFN-γ levels in the spinal cord, astrocytes were converted into antigen-presenting cells. This conversion might promote the progression of pathological damage and result in a peak of EAE severity.

Characterization and primary functional analysis of phenylalanine ammonia-lyase gene from Phyllostachys edulis.

UNLABELLED: Phenylalanine ammonia-lyase (PAL) catalyzes the first reaction in phenylpropanoid pathway leading to the production of phenolic compounds with a wide range of biological functions. The cDNA encoding PAL was isolated from Phyllostachys edulis by reverse transcription-polymerase chain reaction (RT-PCR) and by 5' and 3' rapid amplification of cDNA ends, and was designated as PePAL. The full length of PePAL was 2,503 bp which contained an open reading frame (ORF) encoding a peptide of 701 amino acids, with a theoretic isoelectric point of 6.49 and a calculated molecular mass of 75.7 kDa. PePAL was heterologously expressed in Escherichia coli and the recombinant proteins exhibited both PAL and tyrosine ammonia-lyase (TAL) activities. The optimum temperature and pH of the recombinant PePAL were 50 °C and 8.5-9.0, respectively. The K (m) and V (max) values for L-phenylalanine was calculated as 422 µM and 45.9 nM s⁻¹, while for L-tyrosine were 78 µM and 7.09 nM s⁻¹, respectively. Tissue-specific expression assay showed that PePAL expressed highest in stem and sheath, followed by leaf, and lowest in root. Though the accumulation of PePAL proteins was observed in all these four organs by Western blotting, the highest was detected in leaf. Immunohistochemistry study showed that PePAL was localized primarily in vascular bundles and part of sclerenchyma cells of leaf, sheath and root. These results suggested that PePAL had similar expression pattern and biochemical properties with PALs in other plants, which laid the basis for molecular engineering to improve the quality of bamboo products. KEY MESSAGE: PePAL was a protein with bifunctional enzyme activities of PAL and TAL as shown in vitro assays, and localized primarily in bamboo vascular bundles.

First Report of Medical Tree Peony Root Rot Caused by Fusarium solani in Tongling, China.

Tree peony bark, a main component of Chinese traditional medicine used for alleviating fever and dissipating blood stasis, is mainly produced in Tongling, China. Recently, tree peony cultivation in this area was seriously affected by root rot, with approximately 20 to 30% disease incidence each year. The disease severely affects yield and quality of tree peony bark. During the past 2 years, we collected 56 diseased tree peony plants from Mudan and Fenghuang townships in Tongling. We found reddish brown to dark brown root rot in mature roots, especially on those with injuries. Plant samples collected were disinfected with 2% sodium hypochlorite and isolations were conducted on potato sucrose agar (PSA). Eleven isolates were obtained and all had white fluffy aerial hypha on PSA. Two types of conidia were produced; the larger, reaphook-shaped ones had three to five septa and the smaller, ellipse-shaped ones had one or no septum. The reaphook-shaped conidia were 20.15 to 37.21 × 3.98 to 5.27 µm and the ellipse-shaped conidia were 6.02 to 15.52 × 2.21 to 5.33 µm in size. Chlamydospores were produced, with two to five arranged together. Biological characteristics of the fungi indicated that the optimum temperature for the mycelial growth on PSA was 25 to 30°C and the optimum pH range was 5.5 to 7.0. The above morphological characteristics point the fungal isolates to be Fusarium solani. To confirm pathogenicity, 30 healthy 1-year-old tree peony seedling plants were grown in pots (25 cm in diameter) with sterilized soil and a conidial suspension from one isolate (FH-1, 5 × 105 conidia/ml) was used for soil inoculation. Inoculated seedlings were maintained at 28°C in a greenhouse with a 12-h photoperiod of fluorescent light. Seedlings inoculated with distilled water were used as controls. After 3 weeks, the roots were collected and rinsed with tap water. Dark brown lesions were observed in the inoculated mature roots but not in the control roots. To confirm the identity of the pathogen, F. solani strains were reisolated from the lesions and total genomic DNA was extracted with the cetyltriethylammnonium bromide method from the mycelia of the reisolated strains (1). PCR was performed using the fungal universal primers ITS4 (5'-TCCTCCGCTTATTGATATGC-3') and ITS5 (5'-GGAAGTAAAAGTCGTAACAAGG-3') to amplify a DNA fragment of approximately 590 bp. The purified PCR products were sequenced (Invitrogen Co., Shanghai, China) and shared 100% sequence identity with each other. A comparison of the sequence (JQ658429.1) by the Clustal_W program (2) with those uploaded in GenBank confirmed with the fungus F. solani (100% sequence similarity to isolate S-0900 from the Great Plains of the United States; EU029589.1). To our knowledge, this is the first report of F. solani causing medical tree peony root rot in China. The existence of this pathogen in China may need to be considered for developing effective control strategies. References: (1). C. N. Stewart et al. Biotechniques 14:748, 1993. (2). J. D. Thompson et al. Nucleic Acids Res. 22:4673, 1994.