Secondhand smoke exposure can increase the risk of first ischemic stroke: A 10.7-year prospective cohort study in China.

INTRODUCTION: Passive smoking is considered a major public health issue in China. Prospective evidence regarding the link between secondhand smoke (SHS) and ischemic stroke in China is scarce. METHODS: The China Kadoorie Biobank (CKB) study in Liuzhou City recruited 50,174 participants during 2004-2008. Of these 30,456 never-smokers were included in our study. The median follow-up period was 10.7 years. The incidence of ischemic stroke was obtained through the China Disease Surveillance Points (DSP) system and the Health Insurance (HI) database. Cox proportional risk models were used to evaluate the association between SHS exposure and ischemic stroke. RESULTS: During 320,678 person-years of follow-up, there were 2059 patients with ischemic stroke observed and the incidence of ischemic stroke was 6.42 per thousand person-years. Participants exposed to SHS daily faced a 21 % higher risk of ischemic stroke (HR = 1.21, 95 %CI: 1.09-1.34) compared to those exposed to SHS less than once a week. Subgroup analyses revealed that daily SHS exposure was linked to heightened risk of ischemic stroke among women, non-employed, and non-weekly tea drinkers. CONCLUSIONS: Daily SHS exposure was associated with higher risks of ischemic stroke. Proactive tobacco control strategies are necessary to decrease the risk of ischemic stroke in never smokers.

First Report of Lasiodiplodia hormozganensis Causing Leaf Spot Disease on Areca catechu in China.

The areca palm (Areca catechu L.) is one of the significant cash crops in Hainan Province (China), and a valuable tropical medicinal plant (Cao et al. 2020). In September 2020, spots were found on about 80% of the area of the leaves in a 1,000-acre plantation of areca palms in Haikou City, Hainan Province, and the average incidence was 25%. Initially, Elliptical or irregular dark brown spots appeared on the leaves, with an average size of about 1.5 cm2. With the further expansion of the disease, the spot turned light brown in the center with dark brown edges and a prominent yellow halo. Later stage of the disease, the spots became grayish-white in the center, with obvious whorls, on which many small black spots (pycnidia) were scattered. Eventually the leaves dried out. Ten leaves with typical symptoms were collected from the field. Lesion marginal tissues (5×5 mm2) were surfaced sterilized in 75% ethanol for 20 s, followed by 4 min in 1% NaClO, rinsed 3 times with sterile water, plated on PDA and incubated at 28 ℃. A fungus was isolated with a 98% isolation rate. This strain was named HNAC-5. Subcultures were 80 mm in diameter, white, villous, and neatly edged, after two days of incubation at 28 ℃ in dark. Pycnidia were solitary or clustered in stromata, with orifices that oozed black liquid. Conidiogenous cells were colorless and short cylindrical. Conidia unicellular, initially hyaline, aseptate, ellipsoid to ovoid with granular content, becoming pigmented, 1-septate with longitudinal striations, and measuring 20-31×10-13 µm (n=100). These morphological characteristics were similar to Lasiodiplodia spp. (Abdollahzadeh et al. 2010). The internal transcribed spacer region of rDNA, ß-tubulin gene, and translation elongation gene were amplified using ITS1/ITS4, Bt2a/Bt2b, and EF1-728F/EF1-986R primers, respectively (Alves et al. 2008; Glass and Donaldson 1995; White et al. 1990). The resulting sequences were deposited in GenBank under accession numbers OR272043, OR282568, and OR282567. BLAST analysis showed that the three sequences of HNAC-5 were more than 99% similar to strain CBS 124709 of L. hormozganensis. Phylogenetic analysis was performed using the maximum likelihood method based on the three-gene combined dataset with MEGA 7.0 software. The results indicated that HNAC-5 was grouped in the same clade as other L. hormozganensis Abdollahzadeh, Zare & A.J.L. Phillips. Pathogenicity test was carried out on 15 healthy leaves by in vivo inoculation. Ten leaves were pricked with a sterile needle and divided into group 1 and 2. The remaining five uninjured leaves were group 3. Group 1 and 3 were inoculated with 5-mm-diameter mycelial plugs obtained from 3-day cultures, and group 2 treated with PDA plugs served as controls. Fifteen leaves were cultured at 28°C and 100% relative humidity. After 5 days, leaves of group 1 showed symptoms of the disease and on the tenth day showed the same symptoms as the initial onset of the disease in the field, while leaves of Group 2 and 3 showed no symptoms. Pathogenicity tests were conducted three times with the same results. L. hormozganensis was re-isolated from the inoculated symptomatic leaves, thus, Koch's postulates were confirmed. In China, L. hormozganensis has been reported to cause Bougainvillea spectabilis Willd. branch blight disease (Li et al. 2015), and Scaevola taccada (Gaertn.) Roxb. leaf spot disease (Zhang et al. 2020). To our knowledge, this is the first report of L. hormozganensis causing leaf spot disease on A. catechu in China.

Spicy food consumption reduces the risk of ischaemic stroke: a prospective study.

Previous studies revealed that consuming spicy food reduced mortality from CVD and lowered stroke risk. However, no studies reported the relationship between spicy food consumption, stroke types and dose­response. This study aimed to further explore the association between the frequency of spicy food intake and the risk of stroke in a large prospective cohort study. In this study, 50 174 participants aged 30­79 years were recruited. Spicy food consumption data were collected via a baseline survey questionnaire. Outcomes were incidence of any stroke, ischaemic stroke (IS) and haemorrhagic stroke (HS). Multivariable-adjusted Cox proportional hazard models estimated the association between the consumption of spicy food and incident stroke. Restricted cubic spline analysis was used to examine the dose­response relationship. During the median 10·7-year follow-up, 3967 strokes were recorded, including 3494 IS and 516 HS. Compared with those who never/rarely consumed spicy food, those who consumed spicy food monthly, 1­2 d/week and 3­5 d/week had hazard ratio (HR) of 0·914 (95 % CI 0·841, 0·995), 0·869 (95 % CI 0·758, 0·995) and 0·826 (95 % CI 0·714, 0·956) for overall stroke, respectively. For IS, the corresponding HR) were 0·909 (95 % CI 0·832, 0·994), 0·831 (95 % CI 0·718, 0·962) and 0·813 (95 % CI 0·696, 0·951), respectively. This protective effect showed a U-shaped dose­response relationship. For obese participants, consuming spicy food ≥ 3 d/week was negatively associated with the risk of IS. We found the consumption of spicy food was negatively associated with the risk of IS and had a U-shaped dose­response relationship with risk of IS. Individuals who consumed spicy food 3­5 d/week had a significantly lowest risk of IS.

First Report of Meloidogyne enterolobii Infecting Yellow pitaya (Selenicereus megalanthus) in China.

Yellow pitaya, Selenicereus megalanthus, is a night-blooming, climbing cacti of tropical origin, which has received increasing attention for its potential as a new exotic fruit crop (Lichtenzveig et al. 2000). The crop is grown extensively in Hainan Province, China (3000 ha). In October 2021, a survey was conducted on a farm located in Changjiang (19°21'4″N, 108°47'2″S), Hainan Province, China. Some yellow pitaya plants were found that were stunted and chlorotic, with abnormally thin stems (Fig. 1B), and no symptoms on healthy plants (Fig. 1A). Dead plants were also observed. Many galls and females with egg masses were observed on roots (Figs. 1C & 1D). This is typical of root-knot nematode (RKN) infections, and the incidence of infection was 36.7%. Meloidogyne sp. females and egg masses were dissected from roots of the infected plants. The perineal pattern of females (n= 5) was round to oval-shaped with a high dorsal arch (Figs. 1I & 1J). Second-stage juveniles (J2s) had truncated lips (Figs. 1E & 1F) and long-conical tails with bluntly rounded tips (Figs. 1G & 1H). The J2s body length (n= 24) averaged 416.79 µm (349.21 to 472.76 µm) with a mean width of 15.36 µm (12.47 to 17.52 µm); mean stylet length was 11.16 µm (10.10 to 13.23 µm); tail length averaged 53.73 µm (43.46 to 65.90 µm). The morphological characteristics matched the original description of M. enterolobii (Yang and Eisenback 1983). Males were not found. Genomic DNA was extracted from eight single J2s, and the mitochondrial (mtDNA) region between COII and 16S rRNA gene was amplified with primers C2F3/1108 (Powers and Harris 1993). A 652-bp DNA fragment was obtained, for which the sequence (GenBank accession no. OP122499) was 100% identical to the sequences of M. enterolobii isolates from China（MN269947）and the USA (MN809527). Furthermore, species identification was also confirmed using M. enterolobii specific primers Me-F/Me-R. An amplicon size of ∼230 bp was obtained, which is consistent with those previously reported for M. enterolobii (Fig. 2) (Long et al. 2006). Therefore, this population was identified as M. enterolobii based on morphological and molecular characteristics. Pathogenicity tests were performed in the greenhouse at 26℃ and 80% relative humidity with a 14-h/10-h light/dark photoperiod. Ten RKN-free S. megalanthus seedlings were transplanted into pots containing sterilized soil. After 3 weeks, the roots of 5 plants were inoculated with 3,000 eggs and J2s of M. enterolobii per plant. Five uninoculated plants were used as control plants. After 2 months, no galling or symptoms were observed on the control plants. All inoculated plants had galled roots similar to those observed in the field. Females and egg masses were obtained by dissecting galls. The nematode reproduction factor (RF= final population/initial population) was 1.9. Adult females (n= 5) dissected from inoculated plants were identified as M. enterolobii with sequence-specific primers Me-F/Me-R, thus confirming pathogenicity. The pathogenicity test was carried out twice with similar results. M. enterolobii is one of the most damaging species of RKN, due to its wide host range, high level of pathogenicity, and ability to develop and reproduce on several crops with resistance genes to other RKN (Castagnone-Sereno 2012). To our knowledge, this is the first report of S. megalanthus (yellow pitaya) as a host of M. enterolobii in China. Further studies are needed to develop and evaluate integrated management strategies.

Survival times of HIV/AIDS in different AIDS Diagnostic and Treatment Guidelines from 2006 to 2020 in Liuzhou, China.

BACKGROUND: To compare the survival rates of four timing of treatment initiation for people living with HIV/AIDS provided in China in 2006, 2011, 2015, and 2018, and to investigate the factors impacting survival time. METHODS: A people living with HIV/AIDS retrospective cohort study was in Liuzhou City from April 2006 to December 2020. The information was obtained from the National Comprehensive AIDS Prevention and Control Information System. Life tables and the Kaplan-Meier method were used to calculate participant survival rates and time. The univariate and multivariate Cox regression models were used to investigate the factors related to survival. RESULTS: 18,543 participants were included in this study. In four periods, the 1-year survival rates were 81%, 87%, 95%, and 95%. The 2-year survival rates were 76%, 85%, 93%, and 94%. The 3-year survival rates were 73%, 84%, 92%, and 94%. Results of multivariate Cox regression showed that sex, age of HIV diagnosis, ethnicity, household registration, occupation, marital status, the timing of treatment, education level, route of HIV transmission, whether receiving antiretroviral therapy (ART), and the count of CD4+T cells at baseline (count of CD4+T cells at HIV diagnosis) were factors that are significantly correlated with mortality caused by HIV infection. CONCLUSIONS: With the Guidelines updated from 2006 to 2020, the 1-, 2-, and 3-year survival rates of people living with HIV/AIDS in four periods tended to increase. The timing of treatment initiation of the updated edition of the AIDS Diagnostic and Treatment Guidelines (Guidelines) significantly prolonged the survival time of people living with HIV/AIDS.

Age at smoking initiation and smoking cessation influence the incidence of stroke in China: a 10-year follow-up study.

Our study aimed to explore the correlation between age at smoking initiation and smoking cessation for the risk for stroke in China. We investigated 50,174 participants from one of the urban areas of China Kadoorie Biobank (CKB) Study. Hazard ratios (HRs) and 95% confidence intervals (95% CIs) for association between smoking and incidence of stroke were estimated using Cox regression model. During a median of 10.7 years of follow-up, 4370 total stroke cases were documented. Among men, comparing current smokers to never smokers, the HR of total stroke for current smokers was 1.279 (95% CI, 1.134-1.443) for total stroke. The HRs of total stroke were 1.344 (1.151-1.570) for those started smoking at age less than 20 years, 1.254 (1.090-1.443) for those started smoking at age 20-30 years, and 1.205 (1.012-1.435) for those started smoking at age 30 year and above, with a dose-response relation (P for trend, 0.004). Comparing former smokers to current smokers, in the low pack-year group, those stopped smoking at age less than 65 years had a 18.2% decreased risk for total stroke (0.818; 0.673-0.994). The decreased risk was not found in those stopped smoking at age 65 years and above. Similar results were observed in the high pack-year group. In conclusion, we found that current smokers had a higher stroke risk than never smokers, and the risk increased with a younger age at smoking initiation. Smoking cessation can reduce the risk for stroke, especially could benefit from cessation at a younger age.

Characterization of a novel mycotombus-like virus from the plant-pathogenic fungus Phoma matteucciicola.

Here, we report a novel mycotombus-like mycovirus, tentatively named "Phoma matteucciicola RNA virus 2" (PmRV2), derived from the phytopathogenic fungus Phoma matteucciicola strain HNQH1. The complete PmRV2 genome is comprised of a positive-sense single-stranded RNA (+ssRNA) of 3,460 nucleotides (nt) with a GC content of 56.71%. Sequence analysis of PmRV2 indicated the presence of two noncontiguous open reading frames (ORFs) encoding a hypothetical protein and an RNA-dependent RNA polymerase (RdRp), respectively. PmRV2 contains a metal-binding 'GDN' triplet in motif C of RdRp, while most +ssRNA mycoviruses contained a 'GDD' motif in the same region. A BLASTp search showed that the RdRp amino acid sequence of PmRV2 was most closely related to the RdRp of Macrophomina phaseolina umbra-like virus 1 (50.72% identity) and Erysiphe necator umbra-like virus 2 (EnUlV2, 44.84% identity). Phylogenetic analysis indicated that PmRV2 grouped together with EnUlV2 within the recently proposed family "Mycotombusviridae".

First Report of Colletotrichum fructicola Causing anthracnose on macadamia in China.

Macadamia (Macadamia ternifolia Maiden and Betche) is an important commercial crop in the world and has the reputation of being the king of nuts (Liu et al. 2019). In August 2020, symptoms of anthracnose appeared on leaves of macadamia in Chongzuo, Guangxi Province, China, with an incidence of 15-20%. The disease developed from the edge of leaf. Initially, the disease symptoms on leaves were faded green spots, light yellow. After expanding and linking together, the leaves appeared brown or black irregular spots, and the edges of diseased leaves dried up and formed large necrosis, eventually leading to defoliation. A large number of orange-yellow spots (acervuli) developed on the diseased parts. Under high humidity conditions, the diseased part was grayish-brown or black, and a large number of yellowish-brown conidia were produced on the leaf surface (Fig.1 A-E). Ten symptomatic leaves were collected and washed with distilled water. Twelve lesion marginal tissues were sterilized with 75% ethanol (V/V) for 30 s and 1% NaOCl for 1min and rinsed with sterile distilled water, plated on potato dextrose agar (PDA) and incubated at 28°C under light. After 3 days, the incubated samples all produced similar cultural morphology. One isolate named GXMC2 as a representative was selected for following study. The colony by single-spore purification on PDA were grayish green with a white outer ring and cottony on surface, pale blackish green in reverse side (Fig.1 F). Conidia with oil droplets were solitary, cylindrical, transparent and measuring 13.78 to 19.25 µm (average 16.90 µm) × 5.14 to 7.33 µm (average 6.23 µm) (n=100) (Fig.1 G). Appressoria were brown to dark brown, with different shapes such as ovoid, elliptical or irregular, some with lobes. The average size was 7.89 to 13.25 µm (average 10.64 µm) × 5.76 to 9.02 µm (average 7.86 µm) (n=100) (Fig.1 H). No setae were found. The isolate was identified as Colletotrichum fructicola on the basis of the morphology of the colonies, conidia and appressoria (Park et al. 2018). The six target gene sequences, including internal transcribed spacer (ITS), ß-tubulin (TUB), actin (ACT), histone3 (HIS3), chitin synthase A (CHS), and glyceraldehydes-3-phosphate dehydrogenase (GAPHD) (Qiu et al. 2020), were selected for PCR amplification. The resulting sequences were deposited in GenBank under accession numbers MZ821661, MZ821660, MZ821662, MZ821663, MZ821664 and MZ821665 respectively. Phylogenetic analysis of the concatenated sequences were performed with MEGA 7.0 software. The isolate was grouped in the same clade as other C. fructicola (Fig.2). In May 2022, Inoculation was conducted in the field. Four-year-old macadamia leaves were disinfected with 75% alcohol. The conidial suspension was sprayed on 5 unwounded healthy leaves, and 5 leaves sprayed with sterile distilled water served as control. The experiment was replicated 3 times, with each replicate containing 5 leaves. The average daily temperature and average daily relative humidity in the field were 30°C and 62%, respectively. After 2 days, yellow-brown spots appeared on the inoculated leaves and expanded outward. After 4 days, the diseased areas were dark brown, and the controls remained asymptomatic. The same fungal pathogen was reisolated and purified from inoculated leaves and the identity was confirmed by morphological characterization and molecular analysis, confirming Koch's postulates (Fig.1 I-J). In China, C. fructicola has been reported on Passiflora edulis Sims, Brassica parachinensis, Illicium verum, Peucedanum praeruptorum, etc. (Li et al. 2021; Yu et al. 2022; Zhao et al. 2021; Ma et al. 2020). To our knowledge, this is the first report of anthracnose of macadamia caused by C. fructicola in China. This study provides the basis for further research on this disease.

The Small Secreted Protein FoSsp1 Elicits Plant Defenses and Negatively Regulates Pathogenesis in Fusarium oxysporum f. sp. cubense (Foc4).

Fusarium wilt of banana (Musa spp.), a typical vascular wilt disease caused by the soil-borne fungus, Fusarium oxysporum f. sp. cubense race 4 (Foc4), seriously threatens banana production worldwide. Pathogens, including vascular wilt fungi, secrete small cysteine-rich proteins during colonization. Some of these proteins are required for pathogenicity. In this study, 106 small secretory proteins that contain a classic N-terminal signal peptide were identified using bioinformatic methods in Foc4. Among them, 11 proteins were selected to show transient expressions in tobacco. Interestingly, transient expression of FoSsp1 in tobacco, an uncharacterized protein (of 145 aa), induced necrotic cell death reactive oxygen burst, and callous deposition. Furthermore, the expression of FoSSP1 in Foc4 wild type (WT) was up-regulated during the stage of banana roots colonization. A split-marker approach was used to knock out FoSSP1 in the Foc4 WT strain. Compared with the WT, the deletion mutant Fossp1 was normal in growth rate but increased in conidiation and virulence. RT-qPCR analysis showed that the expression of four conidiation regulator genes in the Fossp1 deletion mutant was significantly decreased compared to the WT strain. In addition, the expression of four pathogenesis-related genes of bananas infected with Fossp1 deletion mutant was down-regulated in comparison with that of the WT. In summary, these results suggested that FoSSP1 is a putative elicitor that negatively regulates conidiation and pathogenicity in Foc4.

A novel hexa-segmented dsRNA mycovirus confers hypovirulence in the phytopathogenic fungus Diaporthe pseudophoenicicola.

A novel hexa-segmented double-stranded RNA (dsRNA) mycovirus was isolated and characterized from the filamentous phytopathogenic fungus Diaporthe pseudophoenicicola and was named Diaporthe pseudophoenicicola chrysovirus 1 (DpCV1). The full-length cDNAs of dsRNA1-6 were 3335, 3030, 3039, 2980, 963, and 780 bp, respectively. Sequence analysis indicated the presence of nine open reading frames (ORFs) in the DpCV1 genome. ORF1 in dsRNA1 putatively encoded the RNA-dependent RNA polymerase (RdRp) and ORF3 in dsRNA2 encoded a capsid protein (CP). The seven remaining ORFs, ORF2 in dsRNA2, ORF4 in dsRNA3, ORF6, seven in dsRNA4, ORF8 in dsRNA5, and ORF9 in dsRNA6, encoded proteins with unknown functions. Phylogenetic analysis revealed that DpCV1 is closely related to members of the cluster I group within the family Chrysoviridae but formed a separate clade. Importantly, all the six segments of DpCV1 were cured successfully through single spore isolation to obtain the isogenic virus-free strains. DpCV1 can confer hypovirulence to the fungal host of Diaporthe pseudophoenicicola. Compared with the virus-free strain, WC02 harbouring the DpCV1 is more sensitive to fungicide prochloraz. Furthermore, the cell wall of DpCV1 infected strain was loose and enlarged. This is the first report of a hexa-segmented tentative chrysovirus in D. pseudophoenicicola.

Discovery of zeylenone from Uvaria grandiflora as a potential botanical fungicide.

BACKGROUND: Botanical pesticides play an important role in organic agricultural practices and are widely used in integrated pest management (IPM). Uvaria grandiflora was mainly reported as traditional medicines and possessed antibacterial, antioxidant, and antiprotozoal activities. Therefore, important biological activities of U. grandiflora may suggest that they have the potential to be used as botanical pesticides. RESULTS: The extract of U. grandiflora exhibited broad-spectrum inhibitory activity toward phytopathogenic fungi and oomycetes, particularly against Colletotrichum musae and Phytophthora capsici, and its secondary metabolite zeylenone also displayed strong antifungal and anti-oomycete activities against phytopathogens. Particularly, half maximal effective concentration (EC50 ) values of zeylenone against Phytophthora capsici and C. musae were 6.98 and 3.37 µg mL-1 , showing better inhibitory effects than those of commercial fungicides (azoxystrobin and osthole). Additionally, the pot experiments showed that the extract of U. grandiflora could effectively control Pseudoperonospora cubensis, Phytophthora infestans, Phytophthora capsici and Podosphaera xanthii. In the field experiment, 5% microemulsion of U. grandiflora extract exhibited 79.72% efficacy against cucumber powdery mildew at 87.5 g ha-1 on the 14th day after two sprayings, which was better than that of 21.5% trifloxystrobin and 21.5% fluopyram SC at 200.9 g ha-1 . Surprisingly, 5% microemulsion of U. grandiflora extract could promote cucumber growth significantly. Furthermore, the action mechanism analysis indicated that zeylenone may damage the cytoderm and affect energy metabolism of Phytophthora capsici. CONCLUSION: It is the first time that the extract of U. grandiflora and zeylenone have been discovered leading to broad application prospects in the development as botanical fungicides. © 2021 Society of Chemical Industry.

Photostable 1-Trifluoromethyl Cinnamyl Alcohol Derivatives Designed as Potential Fungicides and Bactericides.

In the current work, a series of 1-trifluoromethyl cinnamyl alcohol derivatives were designed and synthesized and their antifungal activities were evaluated. The bioassay result showed that most compounds exhibited excellent antifungal activity in vitro at 10 µg mL-1. Next, photostable and easily synthesized compound 2 with broad-spectrum antifungal activity in vitro was selected as a potential candidate to evaluate its antibacterial and antifungal activities. The EC50 values of compound 2 against eight fungal plant pathogens in vitro ranged from 3.806 to 17.981 µg mL-1; at the same time, compound 2 could effectively control Podosphaera xanthii, Odium heveae Steinm, Puccinia striiformis West, and Puccinia sorghi in pot experiments. In addition, compound 2 exhibited excellent antibacterial activities in vitro and in vivo against Xanthomonas oryzae pv. oryzae. Furthermore, the absorption and translocation of compound 2 in wheat plants were determined by the high-performance liquid chromatography method. The result showed that compound 2 could be translocated acropetally as well as basipetally in wheat plants. Finally, it was found that compound 2 had no cross-resistance with carbendazim, azoxystrobin, and boscalid.

Complete genome sequence of a novel mycovirus from Phoma matteucciicola.

The complete genome sequence of a novel mycovirus, Phoma matteucciicola RNA virus 1 (PmRV1), derived from Phoma matteucciicola strain LG-01, was sequenced and analyzed. The complete cDNA sequence of PmRV1 is 3432 bp in length with a GC content of 57.17%. The genome of PmRV1 contains two putative open reading frames (ORFs): ORF1 and ORF2. ORF1 encodes a hypothetical protein with significant similarity to a protein encoded by Periconia macrospinosa ambiguivirus 1 (PmAV1). ORF2 encodes a protein of 491 amino acids with a conserved RNA-dependent RNA polymerase (RdRp) domain. Additionally, the triad within domain III has an asparagine (GDN) instead of the nearly universally conserved aspartic acid (GDD). RdRp phylogeny showed that PmRV1 grouped together with PmAV1 as a sister branch of a new member of the recently proposed family of mycotombus-like viruses. This is first report of the complete sequence of a novel mycovirus, PmRV1, infecting Phoma matteucciicola strain LG-01, the causal agent of leaf blight of Curcuma wenyujin.

First Report of Bipolaris cactivora Causing Flower Rot of Pitaya (Hylocereus costaricensis) in China.

Pitaya (Hylocereus costaricensis), belonging to the Cactaceae family, has rich functional substances, such as a variety of amino acids, which are popular with consumers (Wichienchot et al. 2010). In May 2019, flowers showed symptoms of rot, with an incidence of 15% in a plantation (233.3 ha) in Changjiang (19°46'N; 108°93'E) (Hainan province), China. The initial disease symptoms of flower were small scattered purple-red spot (1~2 mm), including circular, long oval or irregular in shape. The spots were gradually expanded and coalesced, forming abundant reddish-brown lesions. Later, this disease resulted in rotting and blackening of the whole flower. Many black mildew layers (conidiophores and conidia) on the surface of the lesions were observed under compound microscopy. Symptomatic flower tissue (4 cm2) from collecting samples was disinfected in 75% ethanol for 25 s, followed by 1 min in 5% sodium hypochlorite, rinsed 3 times with sterile water, plated on potato dextrose agar (PDA) for 3 days, and incubated at 28ºC. A fungus was consistently isolated from symptomatic flower samples with 90% isolation rate. Resultant colony of the fungus was circular, dark green, velvety, hairy, after 7 days, incubated at 28ºC. Hyphae were septate, 6.2-8.9 µm (average 7.6±0.5) in diameter. Conidia were straight, obclavate, pale to mid brown, 2-6 septate, 23.0 to 42.2 µm (average 31.0±3.2) × 6.5 to 9.8 µm (average 8.0±0.6) (n = 100). The conidia were normally produced germ tubes from one end or both ends. The width of conidiophore was 5.1 to 6.6 µm (average 5.8±0.4) (n = 50). Sequences were generated from the isolate using primers for the internal transcribed spacer region (ITS) (ITS1/ITS4) (White et al. 1990), ribosomal large subunit (LSU) (LROR/LR5) (Vilgalys et al. 1990), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (GPD1/GPD2) (Berbee et al. 1999) loci. The resulting sequences were deposited in GenBank with accession numbers MN960109, MN966852, and MT542865. BLAST analysis demonstrated that these sequences were 99% similar to ITS (HM193535), LSU (MH869295), and GAPDH (HM598681) of Bipolaris cactivora. A maximum likelihood phylogenetic analysis based on combined dataset of ITS, LSU, and GAPDH sequences using MEGA7.0 revealed that the isolate was placed in the same clade as B. cactivora with 100% bootstrap support. A conidial suspension (1 × 105 conidia/ml) of the fungal isolate was prepared by harvesting conidia from pure culture of the fungus grown on PDA 25 days. The 10 mL suspension was sprayed onto ten flowers with no wounding. Ten additional flowers sprayed with sterile distilled water were served as controls. All flowers were covered with plastic bags to maintain high humidity and incubated under natural condition. Typical symptoms of purple-red spot were observed on all the inoculated flowers on the third day. Abundant dark-brown to dark lesions were observed on the surface of flowers and were similar to those observed on the naturally infected flowers after 5 days. The control flowers remained asymptomatic. The fungal isolate of B. cactivora was reisolated from lesion of the flowers and reidentified by morphological and molecular characteristics, thus fulfilled Koch's postulates. Pathogenicity tests were repeated thrice with the same results. B. cactivora had been reported causing flowers and fruit rot of pitaya in South Florida (Tarnowski et al. 2010). This is the first report of B. cactivora causing flower rot of pitaya (H. costaricensis) in China. The flower rot may provide inoculum for the fruit rot, which will cause reduction of pitaya yield.

Molecular Characterization of a Novel Ourmia-Like Virus Infecting Phoma matteucciicola.

Here, we report a novel (+) ssRNA mycovirus, Phoma matteucciicola ourmia-like virus 1 (PmOLV1), isolated from Phoma matteucciicola strain LG915-1. The genome of PmOLV1 was 2603 nucleotides long and contained a single open reading frame (ORF), which could be translated into a product of RNA-dependent RNA polymerase (RdRp) by both standard and mitochondrial genetic codons. Cellular fractionation assay indicated that PmOLV1 RNAs are likely more enriched in mitochondria than in cytoplasm. Phylogenetic analysis indicated that PmOLV1 is a new member of the genus Penoulivirus (recently proposed) within the family Botourmiaviridae.

MicroRNA-1296 expression is associated with prognosis and inhibits cell proliferation and invasion by Wnt signaling in non-small cell lung cancer.

MicroRNAs (miRNAs) are vital regulators of non-small cell lung cancer (NSCLC) development and tumorigenesis. The aim of the present study was to explore the role of miRNA (miR)-1296 expression in NSCLC. The expression of miR-1296 was detected by reverse transcription-quantitative PCR in NSCLC tissues and matched normal tissues. The association of miR-1296 expression with clinicopathological factors of NSCLC patients was evaluated by the χ2 test. Prognostic value of miR-1296 expression levels in patients with NSCLC was assessed using the Kaplan-Meier method and a Cox proportional hazards model; Cell Counting Kit-8, Transwell migration and western blot assays were used to detect the association between miR-1296 and cell proliferation, invasion and Wnt signaling in NSCLC, respectively. The results of the present study demonstrated that miR-1296 expression was significantly downregulated in NSCLC tissues and cells compared to corresponding controls. Lower miR-1296 expression exhibited a significant association with lymph node metastasis and tumor-node-metastasis stage of patients with NSCLC. In addition, the survival analysis demonstrated that low miR-1296 expression predicted a poorer prognosis compared to high miR-1296 expression. Multivariate Cox analysis also demonstrated that reduced miR-1296 expression was an independent risk factor of NSCLC prognosis. Additionally, miR-1296 inhibited cell proliferation, invasion and Wnt signaling in NSCLC. Thus, the results of the present study indicated that miR-1296 expression may be a potential biomarker of NSCLC prognosis and potential target for NSCLC treatment.

The experience of the free superficial palmar branch of the radial artery perforator flap application.

PURPOSE: The purpose of this study is to retrospect and summarize clinical efficacy and experience of the free perforator flap base on the superficial palmar branch of the radial artery for tissue defect reconstruction in hand. METHOD: 17 patients who underwent tissue defect in hands reconstruction by the free superficial palmar branch of the radial artery (SPBRA) perforator flaps in our department from July 2014 to October 2018 were reviewed. RESULTS: All the flaps in our series application were survival uneventful except one, which was necrosis because of venous thrombosis postoperative 5 days, and then the abdominal pedicle flap was executed to recover the defect in second stage. The first dorsal metacarpal artery flap and the arterial venous flap were utilized to cover the defect in one right index finger and one right ring finger due to the absence variation of the SPBRA. 2 cases presented tension vesicle of superficial skin and 1 case occurred venous congestion. All donor sites were closed primarily. The follow-up period means 13.5 months (range, 4-50 months). The static 2 point discrimination test mean 7.53 mm (range, 4-11 mm). All flaps acquire protective feeling at the latest follow-up. The self-assessment of patients: 13 cases in good, 4 cases in fair. CONCLUSION: The goal of physiological reconstruction and esthetic effect can be achieved for hand tissue defect by the free SPBRA perforator flap, multiple tissues of the flap can be contained according to the defect. Even though the SPBRA is variation, arterial venous flap could be applied thanks to abundant superficial cutaneous veins.

Complete genomic sequence and organization of a novel mycovirus from Phoma matteuccicola strain LG915.

The complete genome of a double-stranded RNA (dsRNA) mycovirus, Phoma matteuccicola partitivirus 1 (PmPV1) was sequenced. It consists of two dsRNA segments, 1664 bp (dsRNA-1) and 1383 bp (dsRNA-2) in length, each containing a single open reading frame (ORF) potentially encoding a 46.78-kDa protein and a 40.92-kDa protein, respectively. dsRNA-1 encodes a putative polypeptide with a conserved RNA-dependent RNA polymerase (RdRp) domain that shows sequence similarity to the corresponding proteins of partitiviruses. The protein encoded by dsRNA-2 has no significant similarity to the typical coat proteins (CPs) of partitiviruses, but structure analysis nevertheless suggested that it might function as a coat protein. Purified viral particles of PmPV1 were isometric and approximately 29 nm in diameter. Phylogenetic analysis showed that PmPV1 is closely related to members of the genus Gammapartitivirus within the family Partitiviridae but forms a separate branch with Colletotrichum acutatum RNA virus 1 and Ustilaginoidea virens partitivirus 2. This is the first report of the full-length nucleotide sequence of a novel virus of the genus Gammapartitivirus infecting P. matteuccicola strain LG915, the causal agent of leaf blight of Curcuma wenyujin.

Comparison of immunogenicity between hepatitis B vaccines with different dosages and schedules among healthy young adults in China: A 2-year follow-up study.

Immunogenicity of hepatitis B vaccine between 20 µg with 3-dose schedule and 60 µg with 2-dose regimens was compared 2 years after primary immunization. A total of 353 healthy adults aged 18-25 years were enrolled in the study and randomly assigned (1: 1: 1) into 3 vaccine groups: A (20 µg, 0-1-6 month), B (60 µg, 0-1 month) and C (60 µg, 0-2 month). Serum samples were collected at 1 month after a series vaccination and 12 months, 24 months after the first-dose. The GMC level of anti-HBs antibody was measured using Chemiluminescent Microparticle ImmunoAssay (CMIA). There were 59, 45 and 55 vaccinees available to follow-up with 2 year later in vaccine groups A, B and C, respectively. No significant differences existed in sex ratio, age and body mass index (BMI) among vaccinees at month 24 and the corresponding participants at baseline in each group (P > 0.05). The seroprotection rates in group A, B and C were 98.31%, 88.37% and 85.19%, respectively (P = 0.014), reflecting the fact that the rate of group A was significantly higher than that in group C (P = 0.026). Also, the GMC level of anti-HBs antibody in group A was significantly higher than those of other two groups (427.46 mIU/ml vs. 89.74 mIU/ml, 89.80 mIU/ml, respectively; all P < 0.01). This data suggested that the standard 20 µg (0-1-6 month) regimen of hepatitis B vaccine should be recommended as a priority on the premise of complete compliance in adults.

A novel function of junctional adhesion molecule-C in mediating melanoma cell metastasis.

Hematogenous dissemination of melanoma is a life-threatening complication of this malignant tumor. Here, we identified junctional adhesion molecule-C (JAM-C) as a novel player in melanoma metastasis to the lung. JAM-C expression was identified in human and murine melanoma cell lines, in human malignant melanoma, as well as in metastatic melanoma including melanoma lung metastasis. JAM-C expressed on both murine B16 melanoma cells as well as on endothelial cells promoted the transendothelial migration of the melanoma cells. We generated mice with inactivation of JAM-C. JAM-C(-/-) mice as well as endothelial-specific JAM-C-deficient mice displayed significantly decreased B16 melanoma cell metastasis to the lung, whereas treatment of mice with soluble JAM-C prevented melanoma lung metastasis. Together, JAM-C represents a novel therapeutic target for melanoma metastasis.