[Analysis of the effect of early subcutaneous specific immunotherapy on the levels of dust mite allergen-specific antibodies and polyunsaturated fatty acid metabolism].

Objective: To investigate the effects of subcutaneous immunotherapy (SCIT) on patients' immune markers and metabolic levels in the early stage of allergen treatment, and to gain insight into the role of SCIT in regulating immune responses and metabolic levels, so as to provide reference data for the further discovery of potential biomarkers. Methods: A longitudinal study was used to include 40 subjects who underwent SCIT with dust mite allergens in the Department of Pediatrics of the First Affiliated Hospital of Guangzhou Medical University between November 2017 and February 2022, including 20 subjects each of single mite subcutaneous immunotherapy (SM-SCIT) and double mite subcutaneous immunotherapy (DM-SCIT). In this study, levels of dust mite allergen-specific antibodies and polyunsaturated fatty acid metabolism were measured before and 12 months after treatment, while pulmonary function tests were performed. The therapeutic effects of the patients were followed up by visual analogue scale (VAS), asthma control test (ACT) and total medication scores (TMS). The results were statistically analyzed using t-test and Mann-Whitney U-test. Results: After 12 months of treatment with SCIT, both groups showed a significant decrease in total VAS score (SM-SCIT:Z=-2.298, P<0.05; DM-SCIT:Z=-3.411, P<0.001); total ACT score (SM-SCIT:Z=-2.054, P<0.05; DM-SCIT:Z=-2.014, P<0.05) and total medication scores (SM-SCIT:Z=-3.799, P<0.000 1; DM-SCIT:Z=-3.474, P<0.001) were significantly higher, in addition to significantly higher MMEF75/25 values in the DM-SCIT group (t=-2.253, P<0.05). There was no significant change in sIgE in the SM-SCIT group (P>0.05), and the sIgG4 levels of the Der p, Der f, p 1, p 2, f 2, and p 21 fractions were significantly elevated (Z=-2.651, -3.771, -2.949, -2.912, -2.725, -2.128, and -3.285, respectively, all P<0.05); The sIgE of Der p 2, f 2, p 7 and p 23 fractions(Z=-2.651, -3.771, -2.949, -2.912, -2.725, -2.128, -3.285, all P<0.05) and the sIgG4 levels of the Der p, Der f, p 1, p 2, f 1, f 2, p 10, p 21 and p 23 fractions (Z=-3.808, -3.845, -3.061, -2.688, -2.464, -3.211, -2.371, -2.091, -2.427, all P<0.05) of the DM-SCIT group were significantly elevated. Metabolomics analysis showed that arachidonic acid, docosahexaenoic acid, docosapentaenoic acid, eicosapentaenoic acid, 5, 9, 12-octadecatrienoic acid, 5(S)-hydroxylated eicosatetraenoic acid, and dihomo-gamma-linolenic acid were significantly elevated at the beginning of the treatment period after SM-SCIT treatment (Z of -2.191, -2.497, -1.988, -2.090, -2.19, -2.803, -2.073, all P<0.05); 5(S)-hydroxylated eicosatetraenoic acid showed elevated and alpha-linolenic acid, eicosadienoic acid, and eicosapentaenoic acid were significantly decreased in the DM-SCIT group after treatment (Z=-1.988, -2.090, -2.497, -1.988, respectively, all P<0.05). Correlation analysis showed that arachidonic acid was significantly negatively correlated with changes in dust mite-specific IgG4 (r=-0.499, P<0.05), and that alpha-linolenic acid, 5, 9, 12-octadecatrienoic acid, and eicosapentaenoic acid were positively correlated with the ΔsIgG4 of the dust mite der p 2 (r=0.451, 0.420, 0.474, respectively; all P<0.05). Conclusion: Significant changes in allergen-specific antibody levels and polyunsaturated fatty acid metabolism levels occur during SCIT, and the two may interact and influence each other.

[Pathological classification of malignant peritoneal mesothelioma and comparative analysis with peritoneal carcinomatosis].

Objective: To analyze the pathological classification of malignant peritoneal mesothelioma (MPeM) and screen the immunohistochemical markers that can distinguish MPeM from peritoneal metastatic carcinoma (PC) . Methods: In June 2020, the pathological results of peritoneal biopsy of 158 MPeM and 138 PC patients from Cangzhou Central Hospital, Cangzhou People's Hospital, and Cangzhou Hospital of Integrated Traditional Chinese and Western Medicine from May 2011 to July 2019 were retrospectively analyzed, and the pathological classifications of MPeM in Cangzhou were summarized. Immunohistochemical markers of MPeM and PC patients were analyzed, and receiver operating characteristic curve (ROC curve) was drawn for differential diagnosis of MPeM and PC. Results: There were 55 male and 103 female MPeM patients in Cangzhou, with an average age of 57.1 years old. The asbestos exposure rate was 91.14% (144/158). The most common pathological classifications were cutaneous type, accounting for 90.51% (143/158). There were significant differences in the expression of calreticulum protein, CK5/6, vimentin, D2-40, carcinoembryonic antigen (CEA) and tail type homologous nuclear gene transcription factor 2 (CDX-2) between MPeM and PC (P<0.05). Among the 6 positive markers, the sensitivity of calreticulum protein was the highest (0.905) and CEA was the lowest (0.428) . Conclusion: Calreticulum protein, CK5/6, vimentin, D2-40, CEA and CDX-2 may be used as specific markers to distinguish the diagnosis of MPeM from PC.

Skin Ultrastructure and the Changes of HIF-2α, H-FABP Expression in the Myocardium of Electric Shock Death Rats.

ABSTRACT: Objective To observe the skin ultrastructure change of electric shock death rats and to test the expression changes of hypoxia-inducible factor-2α （HIF-2α） and heart type-fatty acid-binding protein （H-FABP） of myocardial cells, in order to provide basis for forensic identification of electric shock death. Methods The electric shock model of rats was established. The 72 rats were randomly divided into control group, electric shock death group and postmortem electric shock group. Each group was divided into three subgroups, immediate （0 min）, 30 min and 60 min after death. The skin changes of rats were observed by HE staining, the changes of skin ultrastructure were observed by scanning electron microscopy, and the expression of HIF-2α and H-FABP in rats myocardium was tested by immunohistochemical staining. Results The skin in the electric shock death group and postmortem electric shock group had no significant difference through the naked eye or by HE staining. Under the scanning electron microscope, a large number of cellular debris, cells with unclear boundaries, withered cracks, circular or elliptical holes scattered on the cell surface and irregular edges were observed. A large number of spherical foreign body particles were observed. Compared with the control group, the expression of HIF-2α in all electric shock death subgroups increased, reaching the peak immediately after death. In the postmortem electric shock group, HIF-2α expression only increased immediately after death, but was lower than that of electric shock death group （P<0.05）. Compared with the control group, the expression of H-FABP in all subgroups of electric shock death group and postmortem electric shock group significantly decreased. The expression of H-FABP in all subgroups of electric shock death group was lower than that of the postmortem electric shock group （P<0.05）. Conclusion Electric shock can increase HIF-2α expression and decrease H-FABP expression in the myocardium, which may be of forensic significance for the determination of electric shock death and identification of antemortem and postmortem electric shock.

[Application value of non-invasive ventilation combined with high flow nasal cannula oxygen therapy in sequential treatment of patients with chronic obstructive pulmonary disease after mechanical ventilation].

Objective: To investigate the value of non-invasive ventilation (NIV) combined with high flow nasal cannula oxygen therapy (HFNCO) in sequential treatment of patients with chronic obstructive pulmonary disease after mechanical ventilation. Methods: Chronic obstructive pulmonary disease with acute exacerbation (AECOPD) patients with invasive mechanical ventilation (MV) and successful withdrawal admitted into Huxi Affiliated Hospital of Jining Medical College from January 2018 to December 2019 were enrolled for perspective study. The patients were divided into treatment group (n=40) and control group (n=33) by random number table method. The treatment group was given NIV and HFNCO, the control group was given NIV treatment alone. Bedside ultrasound was used to measure the patients' diaphragmatic motion, and the differences between the two groups of patients before treatment, 24, 48 and 72 h after treatment were compared in diaphragmatic excursions during quiet breathing (DEq), diaphragmatic excursions during deep breathing(DEd), diaphragmatic shallow fast breathing index (D-RSBI), arterial oxygen partial pressure (PaO(2)), arterial partial pressure of carbon dioxide (PaCO(2)), re-tracheal intubation rate, mortality rate for 28 days and average duration of NPPV treatment within 3 days. Results: There were no statistically significant differences in DEq, DEd, D-RSBI, PaO(2) and PaCO(2) between the two groups before treatment (all P>0.05). After 24 h treatment, DEd decreased in both groups, D-RSBI increased in both groups, However, D-RSBI [(1.33±0.56) vs (1.62±0.59) times·min(-1)·mm(-1)] in the treatment group was significantly lower than the control group, P=0.034. After 72 h treatment, DEd [(41.4±8.1) vs (37.8±6.0) mm] was significantly higher than the control group, D-RSBI [(1.02±0.27) vs (1.22±0.43) times·min(-1)·mm(-1)] was significantly lower than the control group (all P<0.05). The average duration of NIV treatment time [(7.5±1.2) vs (9.3±2.6) h] in the treatment group was significantly shorter than that in the control group (P<0.01). There were no statistically significant differences in PaO(2), PCO(2), re-tracheal intubation rate and the mortality rate of 28 days. Conclusion: NIV combined with HFNCO sequential therapy can effectively relieve diaphragm fatigue and promote recovery of respiratory muscle strength, and it's better than NIV alone.

[Expression of costimulatory molecules B7H3 and B7H4 in T lymphoblastic lymphoma/leukemia and clinical significance].

Objective: To investigate the expression of B7H3 and B7H4 in T lymphoblastic lymphoma/leukemia (T-LBL/ALL) in correlation with clinicopathological parameters and patient prognosis. Methods: Immunohistochemistry (IHC) was used to detect the expression of B7H3 and B7H4 protein in 100 cases of T-LBL/ALL(test group) and 30 cases of lymph node reactive hyperplasia (LH) (control group), diagnosed at Shanxi Cancer Hospital from January 2001 to June 2017. Real-time RT-PCR was used to detect the mRNA expression of B7H3 and B7H4 in 50 cases of T-LBL/ALL and 30 cases of LH (control group). Results: There were 79 males,21 females. Immunohistochemical results showed that the expression rates of B7H3 and B7H4 were 23%(23/100) and 54%(54/100), respectively. By real-time RT-PCR, the relative expression of B7H3 mRNA in the T-LBL/ALL group was 2.5 times of that of the LH group. The expression levels of B7H4 mRNA in T-LBL/ALL group and LH group were extremely low.Single factor analysis showed that B7H3 protein expression in T-LBL/ALL group was associated with B symptoms and primary nodal disease (P<0.05). B7H4 protein expression was associated with mediastinal broadening and bone marrow involvement (P<0.05). B7H3 protein, B7H3 mRNA, B7H4 protein expression and IPI score were associated with prognosis (P<0.05), and the combined expression of B7H3 and B7H4 was associated with T-LBL/ALL prognosis (P<0.05). Multivariate Cox regression analysis showed that overexpression of B7H3 mRNA was an independent risk factor for the prognosis of patients with T-LBL/ALL (P<0.05). Conclusion: Expression of B7H3 and B7H4 is closely corelated with clinicopathological parameters and prognosis of patients with T-LBL/ALL, suggesting that B7H3 and B7H4 expression play an important role in the development of T-LBL/ALL.

[Effect and significance of mild hypothermia on cerebral blood flow velocity and cerebral extraction rate of oxygen in patients with severe subarachnoid hemorrhage].

Objective: Through studying the variations of cerebral blood flow velocity and cerebral extraction rate of oxygen to investigate the effect and mechanism of mild hypothermia therapy on early brain injury (EBI) and cerebral vasospasm (CVS) induced by sever subarachnoid hemorrhage (SAH). Methods: A total of 62 adult patients admitted in the Department of Neurosurgery of Tianjin TEDA Hospital from January 2014 to December 2016 with severe SAH without contraindications of hypothermia therapy were randomly divided into mild hypothermia (MH) group of 30 cases and routine treatment (RT) group of 32 cases.The general data were no significant difference.The routine treatment group got bloody cerebrospinal fluid drainage, spasmolysis, 3H treatment, etc.Besides conventional treatment, MH group got mild hypothermia therapy started on the day of admission within 2-8 hours, lower rectal temperature and maintained at (35±1) ℃, maintain 5-7 d. The mean velocity of middle cerebral artery blood flow (VmMCA) and Lindergaard index of two groups were detected by transcranial Doppler to indirectly evaluate the degree and evolution of CVS.Blood gas analysis was performed to obtain the blood oxygen content of the artery and jugular vein (CaO(2)/CjvO(2)) in the two groups at the same time, and the cerebral extraction rate of oxygen (CERO(2)) = (CaO(2)-CjvO(2))/CaO(2) was calculated. Results: Within 5 times of admission d1, d2, d3, d7 and d14, mean results of VmMCA of RT group were significantly higher than those of the MH group at d2, d3, d7 and d14 on statistics.The changes of CERO(2) between MH group and RT group during the observation period were compared at the same time: there was no significant difference between d1 and d14 (P>0.05); at d2, d3 and d7 showed marked differences, and that of the MH group was significantly lower than that of the RT group' (P<0.01). The correlation analysis showed that it had a weak correlation between CERO(2) and VmMCA (P>0.05) in the MH group, and CERO(2) was significantly positively correlated with VmMCA in the RT group (P<0.01). Conclusion: MH therapy has a positive significance to reduce the incidence, degree and the duration of CVS.The relationship between CVS and the degree of hypoxia in brain was broken by the MH therapy to reduce the adverse effects of EBI through reducing metabolism, thereby alleviating hypoxia damage in brain tissue.Setting the appropriate target temperature and the course of treatment and then the gentle rewarming process can reduce the incidence of complications of hypothermia therapy.

Estimation on Formation Time of Thrombus.

OBJECTIVES: To observe the changes of the formation time of venous thrombus in rats, and to provide new ideas and methods for the estimation on thrombus formation time of the forensic cases died from thrombosis. METHODS: Totally 80 rats were randomly divided into 10 groups （0 h, 3 h, 6 h, 12 h, 1 d, 3 d, 1 week, 2 weeks, 3 weeks and 4 weeks after operation）. A vein thrombosis model was established by the "narrow" method. The processes of thrombosis, organization, recanalization and the features of change on hemosiderin and calcium salt were observed by HE stain, Perls stain and Von Kossa stain. The expression changes of CD61, α-SMA and CD34 were observed by immunohistochemical staining technique. RESULTS: Platelets adhered to the exposed blood vessel intima 3 h after operation, and platelet trabeculae were formed by the repeated accumulation of platelets 1 d after operation. The thrombus organization formed through the fibroblasts from vessel wall that grew into the interior of the thrombus 3 d after operation. Endothelial cells covered the surface of thrombus and then the new blood vessels were reformed, and the vessels were reconstructed. The expression of CD61 upregulated at the stages of the thrombus formation （3 h） and thrombus reformation （4 weeks）, and reached the peak 1 d after thrombus formation. The release of hemosiderin and the initial expression of α-SMA were detected 3 d later. Calcium deposit and expression of CD34 were observed 1 week later. CONCLUSIONS: The hemosiderin, calcium salt, CD61, α-SMA and CD34 show time-dependent changing characteristics, which is expected to provide a reference for the estimation on thrombus formation time of the forensic cases died from thrombosis.

Development of indicators for evaluating the appropriate use of triazoles for invasive fungal disease: A Delphi panel survey.

WHAT IS KNOWN AND OBJECTIVES: Triazoles including fluconazole, itraconazole, voriconazole and posaconazole are now widely used, whereas in some countries, they are reportedly used in inappropriate way frequently; thus, it is clearly a matter of urgency to regulate the use of triazole drugs. Several studies have made good attempts to evaluate antifungal use, but they did not cover the entire medication process. This study aimed to establish indicators for the appropriate use of triazoles for invasive fungal disease, so as to produce a reference for evaluating and standardizing the rational application of triazole antifungals. METHODS: A comprehensive analysis and literature review were conducted to construct an evaluation system for indications, therapeutic timing, duration of drug usage, drug dosage, administration method, drug interactions, medication in specific populations and pharmaceutical care. A Delphi questionnaire was designed based on the preliminary evaluation of an indicator system that comprised eight primary indicators, 29 secondary indicators and 54 tertiary indicators. A Delphi panel survey was completed by 28 and 26 experts in the first and second rounds, respectively, via e-mail. Agreement was considered to be reached when the median score was in the top tertile (4 or 5), and 80% of panel scores were in the top tertile. The experts mainly comprised physicians and clinical pharmacists from different regions. Based on expert opinions, the evaluation indicator system was modified and the final version was established. RESULTS: The response rate was high in the first and second rounds, at 100% and 93%, respectively, as was the dependability coefficient (0.83). The expert opinions resulted in four secondary indicators and nine tertiary indicators being deleted, and 10 tertiary indicators being modified. After the two rounds of the Delphi survey, the final established system of indicators comprised eight primary indicators, 25 secondary indicators and 45 tertiary indicators. WHAT IS NEW AND CONCLUSIONS: Contrasted with previous studies that have only focused on a subset of indicators, this research establishes comprehensive indicators for evaluating the use of triazoles for invasive fungal disease and which cover most of the medication process: indications, therapeutic timing, duration of drug usage, drug dosage, administration method, drug interactions, medication in specific populations and pharmaceutical care. The indicators can reflect the characteristic of triazoles throughout the process of clinical administration for an invasive fungal disease, and it will be helpful as references when hospital administrators are regulating the use of antifungals.

Resolving the Phoma enigma.

The Didymellaceae was established in 2009 to accommodate Ascochyta, Didymella and Phoma, as well as several related phoma-like genera. The family contains numerous plant pathogenic, saprobic and endophytic species associated with a wide range of hosts. Ascochyta and Phoma are morphologically difficult to distinguish, and species from both genera have in the past been linked to Didymella sexual morphs. The aim of the present study was to clarify the generic delimitation in Didymellaceae by combing multi-locus phylogenetic analyses based on ITS, LSU, rpb2 and tub2, and morphological observations. The resulting phylogenetic tree revealed 17 well-supported monophyletic clades in Didymellaceae, leading to the introduction of nine genera, three species, two nomina nova and 84 combinations. Furthermore, 11 epitypes and seven neotypes were designated to help stabilise the taxonomy and use of names. As a result of these data, Ascochyta, Didymella and Phoma were delineated as three distinct genera, and the generic circumscriptions of Ascochyta, Didymella, Epicoccum and Phoma emended. Furthermore, the genus Microsphaeropsis, which is morphologically distinct from the members of Didymellaceae, grouped basal to the Didymellaceae, for which a new family Microsphaeropsidaceae was introduced.

First Report of Acrostalagmus luteo-albus Causing Red Rust of Needle Mushroom (Flammulina velutipes) in China.

Needle mushroom (Flammulina velutipes) is an edible and medicinal mushroom with annual production yields for China exceeding 2 million metric tons. Hebei province is one of the biggest producers of needle mushrooms in China. From 2009 to 2010, red rust-colored sporulation (fungal mycelia and conidia) was found on the substrate surface of white needle mushroom bags in more than 10 mushroom cultivation operations in Hebei. The rust-colored sporulation covered the substrate surface, where the development of the fruiting bodies was inhibited; the stipes were sparse and became light brown when the substrate was slightly affected. The fruiting bodies on severely affected substrate were unable to complete development or shriveled and died off on the substrate. More than 30 to 40% substrate bags were contaminated with the red rust mold, with approximately a 40% yield reduction in each production house. Single conidia were isolated from the red rust mold and cultured on potato dextrose agar at 25°C in the dark. The colony was round, compact, reddish-brown, and slow-growing (2 to 3 mm/d). Main conidiophore axes were erect, often branched in five to seven layers, tapered from the base to the upper part, pale reddish brown at the base, and almost hyaline at the apex. Two to five phialides usually verticillated at each level along the main stipe and its branches. Phialides were narrowly flask-shaped and only very slightly swollen at the base. Conidia were oval, measured 3.5 to 5.4 (4.5) × 2.3 to 3.6 (3.0) µm, L/W = 1.2 to 2.0 (1.5), and formed pale reddish brown, slimy heads. The isolate was tentatively identified as Acrostalagmus luteo-albus based on its morphological characteristics (2). For molecular analysis, the internal transcribed spacer (ITS) regions of ribosomal DNA from the isolate were PCR-amplified using universal primers ITS1 and ITS4. The resulting sequence (Accession No. KC127681) submitted to GenBank had a 99% identity to that of A. luteo-albus (JN545827) isolated from vineyard soil and Accession No. JQ387575 isolated from twig of declining persimmon tree. To confirm the pathogenicity of the isolate to needle mushroom, five bags of needle mushroom with stipes of 1 to 2 cm long were inoculated with a conidia suspension (105 conidia/ml) of the isolate and incubated at 13 to 15°C in the dark, while five non-inoculated bags (sprayed with sterile water only) were used as a control. Five days after inoculation, the development of the fruiting body was obviously inhibited, and a few growing fruiting bodies became light brown, compared with the non-inoculated fruiting bodies, which were growing healthily and fast and were white with no symptoms. The pathogen was re-isolated from the inoculated fruiting bodies. A. luteo-albus is primarily saprophytic but is sometimes pathogenic. Indeed, it has been associated with post-harvest rot of ginger rhizomes in Brazil (1). To our knowledge, this is the first report of A. luteo-albus causing damage to needle mushroom in China. References: (1) S. I. Moreira et al. Trop. Plant Pathol. 38:218, 2013. (2) R. Zare et al. Mycol. Res. 108:576, 2004.

Evaluation of cellulolytic activity in insect digestive fluids.

Efficient and low-cost cellulolytic enzymes are urgently needed to degrade recalcitrant plant biomass during the industrial production of lignocellulosic biofuels. Here, the cellulolytic activities in the gut fluids of 54 insect species that belong to 7 different taxonomic orders were determined using 2 different substrates, carboxymethyl cellulose (CMC) (approximating endo-ß-1,4-glucanase) and filter paper (FP) (total cellulolytic activities). The use of CMC as the substrate in the zymogram analysis resulted in the detection of distinct cellulolytic protein bands. The cellulolytic activities in the digestive system of all the collected samples were detected using cellulolytic activity analysis. The highest CMC gut fluid activities were found in Coleoptera and Orthoptera, while FP analysis indicated that higher gut fluid activities were found in several species of Coleoptera and Lepidoptera. In most cases, gut fluid activities were higher with CMC than with FP substrate, except for individual Lepidoptera species. Our data indicate that the origin of cellulolytic enzymes probably reflects the phylogenetic relationship and feeding strategies of different insects.

Genetic polymorphisms of the TLR4 gene and their association with susceptibility to type 2 diabetes mellitus in the Chinese population.

Toll-like receptor 4 (TLR4) is potentially an important gene affecting the susceptibility to type 2 diabetes mellitus (T2DM). The objective of this study was to evaluate whether genetic polymorphisms of the TLR4 gene are associated with T2DM susceptibility. This potential association was analyzed in 668 T2DM patients and 672 healthy controls by polymerase chain reaction-restriction fragment length polymorphism and DNA sequencing methods. Two novel genetic polymorphisms (g.12375A>G and g.14367G>A) were investigated, and our data support the idea that the g.14367G>A variant significantly increased susceptibility to T2DM in homozygote comparison (AA vs GG: OR = 2.396, 95%CI = 1.682-3.413, P < 0.0001), heterozygote comparison (GA vs AA: OR = 1.322, 95%CI = 1.050-1.664, P = 0.0175), dominant model (AA/GA vs GG: OR = 1.511, 95%CI = 1.217-1.876, P = 0.0002), recessive model (AA vs GA/GG: OR = 2.093, 95%CI = 1.496-2.927, P < 0.0001), and allele contrast (A vs G: OR = 1.503, 95%CI = 1.279-1.766, P < 0.0001). The allele A of g.14367G>A variants may contribute to the susceptibility to T2DM. However, we failed to detect a similar significantly increased susceptibility to T2DM in the g.12375A>G variant. Our findings suggest that the g.14367G>A genetic polymorphism of the TLR4 gene is associated with the susceptibility to T2DM in the population studied.

First Report of Phytophthora palmivora Causing Fruit Rot of Fig (Ficus carica L.) in China.

Fresh fig (Ficus carica L.) has been grown on a large scale in Beijing, China, since 2011. In late July 2012, a rot disease occurred on immature fruit of fig after a heavy rain (average daily rainfall 170 mm) in Fangshan District, Beijing, which caused about 30% incidence of green fruit on trees. The symptom first appeared as a water-soaked lesion that was covered with a white, fluffy mass of mycelia, followed by a soft, mushy rot of infected area on the fruit. To isolate the causal agent, mycelia and sporangia from 10 symptomatic fruits were suspended in sterile water, spread on potato dextrose agar (PDA) plates, and incubated at 25°C for 18 h. The isolates from each diseased fruit showed the same colonial characteristics. A single sporangium was isolated under a dissecting microscope and transferred onto PDA to obtain a pure culture. On carrot agar, the colony was white and homogeneous with tidy edge, with a few aerial hyphae. Sporangia were obpyriform with obvious papillae and measured 54.7 to 63.8 (59.3) × 26.5 to 36.3 (30.7) µm. The chlamydospores produced in culture were spherical. The pathogen was identified as Phytophthora palmivora based on the morphological characteristics (3) and confirmed with ITS sequences by PCR amplification using rDNA universal primers ITS1 and ITS4. The resulting sequence (Accession No. KC131229) had a 99% identity to that of P. palmivora (JQ354937) isolated from Pachira aquatica. Koch's postulates were conducted by inoculating six surface-sterilized figs with a PDA plug from a 7-day-old culture, with six noninoculated (PDA plugs only) fruits serving as controls. The inoculated fruits were incubated at room temperature in a plastic box covered with film. Symptoms similar to those on the naturally infected fruits began on wounded fruits 48 h after inoculation and on non-wounded fruits 60 h after inoculation, while the six control fruits remained healthy. P. palmivora was reisolated from the symptomatic fruit tissue. P. palmivora is one of the most severe pathogens on edible figs, being reported by Japanese in 1941 (2). Fruit rot of fig caused by the pathogen was reported in Florida in 1984 (1). To our knowledge, this is the first report of P. palmivora leading to fruit rot on fig in China. References: (1) N. E. El-Gholl and S. A. Alfieri, Jr. Proc. Fla. State Hort. Soc. 97:327, 1984. (2) Y. Nisikado et al. Ber. Ohara Inst. 8:427, 1941. (3) Y. N. Yu. Flora Fungorum Sinicorum: Peronosporales (in Chinese) Vol. 6. Science Press, Beijing, 1998.

Role of sphingosine-1-phosphate receptors in the tumor microenvironment: prospects for cancer immunotherapy.

OBJECTIVE: Sphingosine-1-phosphate (S1P) is a sphingolipid protein with anti-apoptotic and pro-survival effects on cancer cells via S1P receptors (S1PRs); however, the role of S1PRs in the tumor microenvironment and immune invasion is still unclear. This study investigated the relationship between S1PR expressions and patient survival and clinical manifestations with respect to the tumor microenvironment and immune infiltration. MATERIALS AND METHODS: The expression levels of five S1PRs were obtained from The Cancer Genome Atlas pan-cancer database and the Kaplan-Meier survival analysis was performed. We predicted the relationship between S1PRs expression levels and patient survival using the univariate Cox proportional hazard regression model. Subsequently, we analyzed correlations between S1PRs expression and infiltrating immune cell subtypes using the Kolmogorov-Smirnov test and the infiltration levels of immune and stromal cells in each tumor using the ESTIMATE algorithm and Spearman's test. RESULTS: The five S1PRs exhibited significant heterogeneity in their expression levels. The expression levels correlated with overall patient survival; however, anti-apoptotic or pro-apoptotic features varied depending on the cancer type. The variable effects of S1PRs on tumors may be related to TGF-ß levels. Our results suggest that S1PRs exert distinct influences on the tumor stem cell index and chemotherapeutic drug sensitivity. CONCLUSIONS: This research provides comprehensive information on the importance of S1PRs in the immune microenvironment, stemness score, sensitivity of human cancer drugs, and cancer prognosis. Interestingly, our findings indicate variations in the expression levels and functions of different S1PR family members. This study highlights S1PRs as potential new targets for antitumor (adjuvant) therapy.

Integrative analyses of radiation-related genes and biomarkers associated with breast cancer.

OBJECTIVE: In addition to significantly reducing breast cancer recurrence risk, radiotherapy also prolongs patients' lives. However, radiotherapy-related genes and biomarkers still remain poorly understood. The present study aimed to identify radiation-associated genes in breast cancer. MATERIALS AND METHODS: Breast cancer data were downloaded from Gene Expression Omnibus (GEO) and UCSC Xena database. The gene ontology (GO) enrichment and gene set enrichment analysis (GSEA) were performed for annotation and integrated discovery. Protein-protein interaction (PPI) network was constructed by STRING database and hub genes were identified. Then, immunohistochemistry and tissue expression of key genes was analyzed by using the Human Protein Atlas (HPA) and GEPIA database. Genes associated with prognosis were identified by performing univariate cox analysis. RESULTS: We identified 341 differentially expressed genes related to radiotherapy in breast cancer patients. PPI analysis revealed a total of 129 nodes and 516 interactions and identified five hub genes (EGFR, FOS, ESR1, JUN, and IL6). In addition, 11 SDEGs THBS1, SERPINA11, NFIL3, METTL7A, KCTD12, HSPA6, EGR1, DDIT4, CCDC3, C11orf96, and BCL2A1 candidate genes can be used as potential diagnostic markers. The calibration curve and ROC indicate good probability consistencies of 3-years and 5-year survival rates of patients between estimation and observation. CONCLUSIONS: Our findings provide novel insight into the functional characteristics of breast cancer through integrative analysis of GEO data and suggest potential biomarkers and therapeutic targets for breast cancer.

DNA polymorphisms of the Hu sheep melanocortin-4 receptor gene associated with birth weight and 45-day weaning weight.

The melanocortin-4 receptor (MC4R) has important roles in regulating food intake, energy balance, and body weight in mammals. In pigs and cattle, MC4R mutations have been identified as genetic markers for growth and traits. Compared with abundant research conducted on other livestock species, little is known about mutations of the ovine MC4R gene. We investigated the effect of MC4R polymorphisms on birth weight and on 45-day weaning weight in 144 Hu sheep. Four single nucleotide polymorphisms (SNPs; g.1016 G/A, g.1240 T/C, g.1264 G/A, and g.1325 A/G) were identified in the 3ꞌ-untranslated region of Hu sheep MC4R by PCR-single-strand conformation polymorphism and DNA sequencing. A haplotype block, containing g.1240 T/C, g.1264 G/A, and g.1325 A/G, was constructed within the Hu sheep MC4R gene. Four SNPs were found to be significantly associated with 45-day weaning weight, while the haplotype block was significantly associated with birth weight. Hu sheep with the genotypes GG in g.1016 G/A or with the genotype CCAAGG in the haplotype block, had higher 45-day weaning weights. We conclude that these 4 SNPs of the MC4R gene have potential as genetic markers for early growth traits in Hu sheep.

First Report of Damping-off Caused by Alternaria japonica on Chinese Cabbage Seedlings in China.

In May 2011, samples of Chinese cabbage (Brassica campestris L. subsp. chinensis Markino) seedlings at the two-to four-leaf stage with damping-off symptoms were collected from greenhouses in the Tibet Autonomous Region of China. Infected stems of the seedlings were constricted at or near the soil surface. On diseased stems, a light to dark brown coloration was demarcated from healthy tissue. Damping-off and death of seedlings occurred as the lesions enlarged, resulting in a significant reduction of seedlings. Diseased stems were cut into 3-mm-long segments, surface-sterilized with 3.5% sodium hypochlorite for 1 min, and rinsed in sterilized water three times before being placed on water agar. A fungus frequently isolated from diseased plants was hyphal tipped under a dissecting microscope and transferred onto potato dextrose agar (PDA) to obtain a pure culture. The isolate grew slowly on PDA at 25°C with a 12-h photoperiod. The colony was white at first and gradually turned gray or grayish-green. No conidia or chlamydospores developed. However, conidia were produced on potato-carrot agar. Conidia were yellow-brown, obpyriform or obpyriform with beaks, had two to eight transverse septa, one to three vertical or oblique septa, and were produced solitarily or often in chains of two or three and measured 43.5 to 85.2 × 21.5 to 28.0 µm. A few conidia without beaks were also present and were nearly round and slightly smaller than the conidia with beaks. Several adjacent chlamydospores with thickened walls were often intercalary. The isolate was tentatively identified as Alternaria japonica based on its morphological characteristics (1,2). For molecular analyses, the internal transcribed spacer (ITS) regions of ribosomal DNA from the isolate were amplified with universal primers ITS1 and ITS4. The resulting sequence (Accession No. JN654465) submitted to GenBank had a 99% identity to that of A. japonica (Accession No. AY154703.1) isolated from leaves of Raphanus sativus. To confirm the pathogenicity of A. japonica, nine healthy 10-day-old Chinese cabbage seedlings were inoculated at the stem base with one PDA plug from a 6-day-old culture, with nine noninoculated (PDA plus only) seedlings serving as controls. Two days after inoculation, symptoms similar to those on the naturally infected plants developed on the inoculated seedlings. No symptoms developed on the controls. The pathogen was reisolated from the stems of inoculated and diseased seedlings. To our knowledge, this is the first report of A. japonica leading to damping off on Chinese cabbage seedlings in China. References: (1) M. P. Corlett and M. E. Corlett. Can. J. Plant Pathol. 21:298, 1999. (2) T. Y. Zhang. Flora Fungorum Sinicorum: Alternaria (in Chinese) Vol. 16. Science Press, Beijing, 2003.